Latest revision as of 20:55, 19 October 2016

Notebook

Protocols

Notebook

date		activity	summary
04-21-16	Bianca	preparation of working area	pipets and other stuff autoclaved
04-28-16	Janis, Bianca	testing of pipets	pipets work well
		assembly of TALEs	we cloned three different constructs in TALEs and transformed them into E.coli: 1) R-SCTALEN left HA Tag 2) R-TALEN left HA Tag 3) R-TALEN right myc Tag
		isolation of plasmids	We isolated the two plasmids 1) iGEM20? and 2) pET 32a+? out of E.coli with a QIAprep(R) Spin Miniprep Kit. The concentrations were 1) 354 ng/ µl and 2) 43.5 ng/ µl dsDNA measured with a NanoPhotometer
05-03-16	Janis	assembly of TALEs, 2nd try	Cells could not be transformed, propably GGC of vectors did not work. Did the assembly again, this time over night.
		PCR/Gels/Prep/Cloning	Primers arrived. Did PCR with N454-F/R from N454-HA and TRX_mod-F/R from PET32. PCR products cloned into PUC57-vector
05-23-16	Bianca	plasmid isolation	Plasmid isolation with QIAprep Spin Miniprep kit. The protocol attached to the kit was used without step 7.
			Plasmids: TRXmodule1 (1), TRXmodule2 (2), iGEMN454-1 (3), iGEMN454-2 (4), 1-1, 1-2, 2-2, 3-1, 3-2, 5 FOK DS wos (without stop), 6 FOK RR wos, 7 Sc FOK wos
		control digest	Control digest with plasmids (5) - (9). Mastermix for (5)-(9): 3 µl EcoRI-HF, 3 µl BamHI-HF, 12 µl buffer CutSmart and 66 µl H₂O, 14 µl of the Mastermix were added to 6 µl of each plasmid DNA
			Control digest with plasmids (1) - (4). Mastermix for (1)-(4): 5 µl BsaI 10 µl CutSmart, 55 µl H₂2O, 14 µl of the Mastermix were added to 6 µl of plasmid DNA
			The digests ran for 1 h at 37 °C
		agarose gel	After the digest 5 µl loadingbuffer was added to the control digest, 20 µl were loaded on a 1 % agarose gel for 30 min and at 140 V. 7 µl of a 1 kb DNA ladder were used.
05-25-16	Janis	sequencing	Plasmids and primer were pipetted.
05-26-16	Bianca	TNF reaction	500 ng DNA, 1 µl Met, ad 50 µl H₂O. Used DNA: iGEM 1-2 (A and B), MPAx SCL, 90 min at 30 °C
		in-vitro-cleavage assay	TC(-): 12,5 µl DNA, 2 µl enzyme, 2 µl NEB3 buffer, 3,5 µl H₂O (+) Ax7: 10 µl DNA, 2 µl enzyme, 2 µl NEB3 buffer, 6 µl H₂O; 30 min at 37 °C
			MiniElute PCR Purification Kit, 100 µl buffer PB, probes frozen with glycerin
05-30-16	Bianca	in-vitro-cleavage assay	2 µl DNA ((+)AX7 und TC(-)), 3 µl NEB3 buffer, 5 µl BSA, 4 µl TNT(-)/Scl + 16 µl H₂O or 13 µl TNT(+) + 7 µl HH₂OO, ad 30 µl HH₂OO
			centrifugation for: 1 h at 37 °C, 20 min at 65 °C, 10 min at 7000xg, 3 min at 14000xg, 20 µl supernatant + 5 µl LB
		agarose gel	1 % agarose gel, 50 ml liquide agarose + 5 µl RedSafe, 20 min at 140 V
		Western blot/preparing probes	at 37 °C, 30- (4 µl) and 30+ (13 µl) with 60 µl Laemmli, after 0, 30, 90 and 180 min. 10 min at 100 °C, stored on ice
		repetition of in-vitro-cleavage assay	SCL/TNT(+) with Ax7
			25 µl TNT+, 4 µl buffer, 4 µl H₂O, ad 40 µl H₂O
			4 µl Scl, 4 µl buffer, 25 µl H₂O, ad 40 µl H₂O
			37 °C
06-01-16	Bianca	SDS gel	15 µl probes for westernblot load on SDS gel (probes from 0, 30, 90 and 180 min + and -),40 min at 200 V
		westernblot	90 min at 120 mA
			membran swing for 1 h in milkpowder and TBST, 1 x in TBST washed, 1 x 5 min in TBST washed, 1 x 10 min in TBST washed
			membran with TBST and α-His (antibody) over night (15 µl antibody in TBST (100 µg/500 µl))
06-02-16	Bianca, Janis		membran washed, second antibody
06-03-16	Janis	transformation	chemically competent E.Coli cells were used, for Ax7L, Ax7R, scAx7L, 1-2; 2 µl DNA used and 50 µl competent cells; 20 min on ice, 45 sec at 42 °C, 1 h at 37 °C in 1 ml LB without antibiotic.
			cells with Ax7L, Ax7R, scAx7L plated on LB and ampicillin. Cells with 1-2 on LB with chloramphenicol. Stored at 37 °C over night.
06-07-16	Wiebke, Theresa, Sabine	Interlab study	vector transformed into Top10 E.coli via heatshock transfomation. 1 h at 37 °C
06-27-16	Bianca	assembly of TALENs	new vector: p3H001; 4 rudiments with 1) Ax7L-DS, 2) Ax7R-RR, 3) Ax7L-scFOK, 4) Ax7R-scFOK
			p3H001 77,0 ng/µl, 2 µl of C-terminal and 2 µl of N-terminal module, 5 µl H₂O, used PCR programm (see protocol)
		transformation	transformation into E.coli with electroporation. 200 µl LB and 30 min at 37 °C. Plated on LB chloramphenicol, stored at 37 °C over night.
06-28-16	Bianca	overnight cultures	5 ml LB and 34 mg/ml chloramphenicol with 1 colony from the plates. 3 cultures picked from each plate. Incubate at 37 °C over night.
06-30-16	Bianca, Kim	plasmid isolation	plasmid isolation with QIAprep Spin Miniprep kit. The protocol attached to the kit was used without step 7. Stored at -20 °C
		overnight cultures	3 colonies from each plate picked, overnight cultures with 5 ml LB, 34 mg/ml chloramphenicol.
07-01-16	Kim	control digest	control digest with the isolated plasmids from 06-30-16. 10 µl plasmid DNA, 2 µl 10x reaction buffer, 0,5 µl EcoRI, 0,5 µl PstI
			1 h at 37 °C and 10 min at 65 °C
		agarose gel	loaded with 15 µl probes from control digest and 7 µl 1 kb plus DNA ladder
		plasmid isolation	used the overnightcultures from 06-30-16; with QIA Prep Spin Miniprep Kit (without step 7)
07-19-16	Janis	retransformation	plasmids 1.1, 2.1, 3.1, 3.2, 4.1 in Top10 E.coli. Incubation in LB with chloramphenicol (1.1: Ax7L-DS, 2.1: Ax7R-RR, 3.1: Ax7L-scFOK, 3.2: Ax7L-scFOK, 4.1: Ax7R-scFOK)
07-20-16	Kim	plasmid isolation	used the cultures from 07-19-16, with QIA Prep Spin Miniprep Kit (without step 7)
07-21-16	Janis	control digest	used plasmids from 06-20-16, with 4 µl plasmid DNA, 0,5 µl XbaI and 0,5 µl PstI, 2 µl buffer and 10 µl H₂O. For 1 h at 37 °C and 10 min at 65 °C.
07-28-16	Sabine, Wiebke, Theresa	PCR	run a PCR with 10 µl 5x HF Phusion Buffer, 5 µl dNTPs, 2,5 µl forward Primer, 2,5 µl reverse Primer, 1 µl template TCR5 H2, 0,5 µl Phusion polymerase, 28,5 µl H₂O
			1. programm: 98 °C 2 min at the beginnig, denaturation at 98 °C for 10 sec, annealing at 60 °C for 10 sec, extension at 72 °C for 30 sec. steps repeated for 35 times. at the and 72 °C for 5 min.
			2. programm: same as the first programm, just the annealing temperature was 66 °C.
		agarose gel	probes loaded on 1 % agarose gel, run for 120 V for 20 min, 1 kb plus DNA ladder.
		transformation	1.1, 2.1, 3.1, 3.2 and 4.1 in BL21DE3 E.coli with electroporation. 1 h at 37 °C , plated on LB with chloramphenicol, incubation at 37 °C
			puc57 in Top10 with heatshock at 42 °C for 40 sec, 200 µl LB added, 1 h at 37 °C. Plated on LB
		gel elution	cut the fragment out of the gel. add 300 µl QG buffer, 10 min at 50 °C, add 100 µl Isopropanol, transfered to column, 1 min at max. rpm, add 400 µl QG buffer, 1 min at max. rpm, add 700 µl PE buffer,
			1 min at 1500 rpm, centrifuge to dry the column, 1 min at max rpm, add 15 µl EB buffer or H₂O, incubate for one minute, centrifugation for 1 min at max rpm.
		cut and ligation	2 µl cut smart buffer, 2 µl ATP, 2 µl puc57, 1 µl SmaI, 1 µl T4 Ligase, 15 µl Elution, 2 µl H₂O, for 2 h at 30 °C
		PCR	repeat the PCR, but used 1 min elonation time at 72 °C
		agarose gel	load gel with PCR probes, run at 120 V for 20 min
08-01-16	Wiebke, Sabine	Plasmid isolation	Plasmid isolation from overnight cultures with iGem Nterm and GFP puc57 (1, 2, 3, 4)
			Used QIA Prep Spin Miniprep Kit without step 7. 4 ml Overnightculture were used.
		control digest	with the isolated plasmids. used 0,5 µl BsaI, 7 µl DNA, 2 µl CutSmart buffer, ad 20 µl (10.5 µl H₂O)
			1 h at 37 °C, 10 min at 65 °C.
		agarose gel	1 % agarose gel, used 7 µl 1 kb DNA ladder, 20 µl from Controll digest with 6 µl 5x LD buffer (20 µl loaded on agarose gel)
			Gel at 120 V for 25 min
08-02-16	Wiebke, Sabine, Louise	plasmid isolation	Repeat the plasmid isolation from 08-01-16
		control digest	with the isolated plasmids. used 1 µl BsaI, 26 µl DNA, 3 µl CutSmart buffer
		agarose gel	1 % agarose gel, used 7 µl 1 kb DNA ladder, 20 µl from control digest with 6 µl 5x LD buffer (20 µl loaded on agarose gel)
08-03-16	Janis
		cloning	GFP - NTH3 Fragment into puc57: 5 µl DNA, 1 µl SmaI, 1 µl T4 Ligase, 1 µl Puc57, 1 µl ATP, 1 µl tango buffer; 1 h at room temperatur (RT)
		transformation	puc57 with GFP - NTH3 in E.coli (Top10)
		transformation	transformation into E.coli (BL21DE3) with electroporation. 200 µl LB and 30 min at 37 °C. Plated on LB chloramphenicol, stored at 37 °C over night.
			Used the DNA 1.1, 2.1, 3.1, 3.2 and 4.1
08-10-16	Kim	overnightcultures	used 4 colonies from 08-03-16 (puc57 with GFP-NTH3 in Top10), 5 ml LB, 5 µl carbenicillin.
08-11-16	Kim	plasmid isolation	plasmid isolation with the overnightcultures from 08-10-16, used 4 ml overnightculture and QIA Prep Spin Miniprep Kit without step 7.
08-12-16	Kim	control digest	plasmids from 08-11-16; 5 µl DNA, 2 µl CutSmart buffer, 0.5 BsaI, 12.5 µl E.coli ; 1 h at 37 °C, 10 min at 65 °C
		agarose gel	1 % agarose gel loaded with 20 µl probe (20 µl control digest and 4 µl 6x loading buffer), 7 µl 1 kb DNA ladder; 27 min at 100 V
08-16-16	Theresa, Lousie, Severin	PCR	like the PCR from 07-28-16
		agarose gel	1 % agarose gel loaded with 60 µl probe (50 µl PCR probe and 12,5 5x loading buffer), 8 µl DNA ladder; 20 min at 120 V
		gel extraction	gel extraction with MiniElute Extraction Kit
		cloning	like the cloning from 08-03-16, but CurSmart buffer was used
		transformation	10 µl cloning stuff, 50 µl competent E.coli cells; add 200 µl LB for 30 min
			10 µl puc57, 50 µl competent E.coli cells; add 200 µl LB for 30 min
08-17-16	Janis	overnightcultures	5 colonies (1-5) picked from transformation (08-16-16), in 5 ml LB with 5 µl Carbenicellin
08-18-16	Janis	plasmid isolation	plasmid isolation (overnight cultures used from 08-17-16) with QIA Prep Spin Miniprep Kit without step 7.
		control digest	control digest with the isolated plasmids (06-18-16), same control digest like the digest from 08-12-16 (5 µl DNA, 2 µl CutSmart buffer, 0.5 BsaI, 12.5 µl H₂O; 1 h at 37 °C, 10 min at 65 °C)
		assembly	assembly oft GFP-NTH3 into iGem vector; 2 µl iGem vector (50 ng/µl), 1 µl hexa-repeat LA, 1 µl hexa-repeat AB, 1 µl hexa-repeat BR (each 50 ng/µl), 2 µl N-terminal module (100 ng/µl), 2 µl C-terminal module (100 ng/µl), 2 µl NEB buffer,
			1 µl enzym Mix (2 µl ATP, 2 µl NEB CutSmart, 1 µl T4-DNA Ligase, 1 µl BsaI) and 7 µl H₂O
			assembly program: 10 min at 40 °C, 10 min at 16 °C (3 cycles), 20 min at 50 °C, 20 min at 80 °C (Final)
		transformation	15 µl assembly, 50 µl competent cells, transformation with electroporation, 200 µl LB, 1 h at 37 °C, plated on LB with chloramphenicol
08-22-16	Kim	overnightcultures	picked 6 colonies from 08-18-16, with LB and Chloramphenicol
08-23-16	Kim	plasmid isolation	plasmid isolation (used 4 ml overnightcultures from 08-22-16) with QIA Prep Spin Miniprep Kit without step 7
		control digest	with plasmids (08-23-16), used 7 µl DNA, 2 µl 10x reaction buffer, 0.5 µl XbaI, 0.5 µl PstI, 10 µl H₂O
			1 h at 37 °C, 10 min at 65 °C
		agarose gel	1 % agarose gel, loaded with 20 µl probes (20 µl control digest and 4 µl 6x loading buffer), 29 min at 100 V
08-24-16	Janis, Kim	transformation	plasmids 1.1, 2.1, 3.1, 3.2, 4.1 in BL21 E.coli., incubation on LB with Chloramphenicol at 37 °C
		control digest	repeat control digest from 08-23-16 with 10 µl DNA (2 µl 10x reaction buffer, 0.5 µl XbaI, 0.5 µl PstI, 7 µl H₂O); 1 h at 37 °C, 10 min at 65 °C
		agarose gel	1 % agarose gel, loaded with 20 µl probes (20 µl control digest and 4 µl 6x loading buffer), 29 min at 100 V
		overnightculures	picked 4 colonies from 08-18-16, with LB and Chloramphenicol
08-25-16	Janis, Kim	plasmid isolation	plasmid isolation (overnightcultures used from 08-24-16 7-10(GFP-NTH3 in iGem vector)) with QIA Prep Spin Miniprep Kit without step 7.
			plasmid isolation (overnightcultures interlab study + control 1, + control 2, - control 1, - control 2)
		transformation	retransformation plasmids 1.1, 2.1, 3.1, 3.2 and 4.1 in Top 10 E.coli, 50 µl competent cells, 20 min on ice, 45 sec at 42 °C, 1 h at 37 °C in 200 µl LB, plated on LB with chloramphenicol
			transformation plasmids for interlab study in Top10 E.coli, 50 µl competent cells, 20 min on ice, 45 sec at 42 °C, 1h at 37 °C in 200 µl LB, plated on LB
08-26-16	Kim	control digest	with plasmids (25-08-16), used 11 µl DNA, 2 µl 10x reaction buffer, 0.5 µl XbaI, 0.5 µl PstI, 6 µl H₂O - different XbaI and PstI stocks used, also repeat control digest with plasmids 1-6
			1 h at 37 °C, 10 min at 65 °C
		agarose gel	1 % agarose gel, loaded with 20 µl probes (20 µl control digest and 4 µl 6x loading buffer), 29 min at 100 V
08-29-16	Bianca	Interlab study, transformation	DNA from Göttingen in 10 µl H₂O; 50 µl competent cells (Top10 E.coli), 2 µl DNA, 20 min on ice, 45 sec at 42 °C, 1 ml LB 1 h at 37 °C, plated on LB with chloramphenicol, incubation at 37 °C
		overnightcultures	3 colonies each plate (from 08-25-16, Top with 1.1, 2.1, 3.1, 4.1), with 5 ml LB and Chloramphenicol, incubate at 37 °C
			3 colonies each plate (from 08-24-16, BL21 with 1.1, 2.1, 3.1, 3.2, 4.1) with 5 ml LB and Chloramphenicol, incubate at 37 °C
08-30-16	Bianca, Theresa	expression	2.5 ml from each overnightculture (from 08-29-16, BL21 with 1.1, 2.1, 3.1, 3.2, 4.1) in 10 ml LB
			Incubation, optical density after 60 min: 1.1 - 0.136, 2.1 - 0.122, 3.1 - 0.181, 3.2 - 0.153, 4.1 - 0.134
			optical density after 115 min: 1.1 - 0.489, 2.1 - 0.494, 3.1 - 0.596, 3.2 - 0.520, 4.1 - 0.420
			optical density after 150 min: 1.1 - 0.728, 2.1 - 0.540, 3.1 - 0.921, 3.2 - 0.792, 4.1 - 0.660
			1 ml, centrifuged, pellet stored at -20 °C (pre induction)
			add 238 µl IPTG, incubate for 1 h and 40 min at 37 °C, 1 ml centrifuged, pellet stored at -20 °C (after induction)
			rest centrifuged, into liquid nitrogen and stored at -80 °C
		plasmid isolation	plasmid isolation (overnightcultures re-transformation from 08-29-16) with QIA Prep Spin Miniprep Kit without step 7
		overnightcultures	100 µl overnightculture (from 08-26-16 interlab study) with 8 ml LB and chloramphenicol, incubation at 37 °C
08-31-16	Bianca, Theresa, Janis	assembly	with 1 µl GFP-NTH3, 1 µl vector (50 ng/µl), 1 µl HexaI (Repeats 1-6), 1 µl HexaII (Repeats 7-11,5), 1 µl T4-Ligase, 1 µl BsaI, 2 µl 10mM ATP, 2 µl CutSmart buffer, ad 20 µl H₂O
			two different vectors were used: iGem vector (with c-terminus c-63) and psaE6 (with c-terminus c-17) and 4 different Hexa combinations each vector
			A: iGem vector and Hexa 42 + HDHx
			B: iGem vector and Hexa 42 + 2xNG
			C: iGem vector and Hexa 42 + 2xNN
			D: iGem vector and pJet Hexa3 rep
			E: psaE6 and Hexa 42 + HDHx
			F: psaE6 and Hexa 42 + 2xNG
			G: psaE6 and Hexa 42 + 2xNN
			H: psaE6 and pJet Hexa3 rep
		interlab study	culture and measurment
09-01-16	Janis, Theresa	transformation	15 µl assambly product, 50 µl competent cells (E.coli, Top10), 20 min on ice, 45 sec at 42 °C, add 200 µl LB, 1 h at 37 °C, plated on LB with Chloramphenicol
		SDS gel	prepared 4 gels
		control digest	with plasmids (08-25-16re-transformation), used 11 µl DNA, 2 µl 10x reaction buffer, 0.5 µl XbaI, 0.5 µl PstI, 6 µl H₂O
			1 h at 37 °C, 10 min at 65 °C
09-05-16	Theresa, Kim	SDS gel	prepared 8 gels
			separating gel (10 %): 27.25 ml H₂O, 33.25 Acrylamid (30 %), 25 ml Tris (1.5 M; pH 8.8), 1 ml APS (10 %), 1 ml SDS (10 %), 40 µl TEMED
			stacking gel: 34 ml H₂O, 8.5 ml Acrylamid (30 %), 6.25 ml Tris (0.5 M; pH 6.8), 0.5 ml APS (10 %), 0.5 ml SDS (10 %), 50 µl TEMED
			first separating gel, than stacking gel; ~1 ml Isopropanol on separating gel, when it is in the chamber (remove it before filling stacking gel in the chamber)
			1. H₂O, 2. Tris, 3. SDS, 4. Acrylamid, 5. APS, 6. TEMED; APS and TEMED everytime at last! After adding them be fast
		overnightcultures	with colonies from tranformation 09-01-16, 4 colonies of the plates A-D, there was nothing on the plates E-H; 5 ml LB, 5 µl Chloramphenicol and the picked colonie
09-06-16	Theresa, Kim	plasmid isolation	plasmid isolation (overnightcultures from 09-05-16, A1-3, B1-3, C1-3, D1-3) with QIA Prep Spin Miniprep Kit without step 7
		transformation	used plasmids from assembly F, G and H (08-31-16), 10 µl assambly product, 50 µl competent cells (E.coli, Top10), 20 min on ice, 45 sec at 42 °C, add 200 µl LB, 30 min at 37 °C, plated on LB with Chloramphenicol
09-07-16	Theresa, Bianca, Kim	control digest	with plasmids (08-25-16), used 4 µl DNA, 2 µl 10x reaction buffer, 0.5 µl XbaI, 0.5 µl PstI, 13 µl H₂O
			1 h at 37 °C, 10 min at 65 °C
		agarose gel	1 % agarose gel, loaded with 20 µl probes (20 µl control digest and 4 µl 6x loading buffer), 29 min at 100 V
		SDS gel	prepared probes: took a little bit of the pellets (from the expression on 08-30-16), also the pre-induction and the after-induction pellets; 80 µl Leammli, resuspent, 10 min at 100 °C
			load gel with 15 µl of the prepared probes, 5 µl Page Ruler best; run for 42 min; gel in coomassi staining solution
		transformation	15 µl plasmid DNA (from plasmid isolation 09-06-16, plasmids A3, A4, B1, B2, C2, D1, D2) 50 µl competent cells (E.coli, BL21), 20 min on ice, 45 sec at 42 °C, add 200 µl LB, 35 min at 37 °C, 100 µl plated on LB with Chloramphenicol
09-08-16	Theresa, Kim	SDS gel	gel 3 h in H₂O and 4 h in destaining solution, after that over night in PBS buffer
		transformation	repeat transformation from 08-07-16, nothing changed
09-09-16	Kim	overnightcultures	used plates from transformation (09-07-16), each plate (A3, A4, B1, B2, D2) one colonie picked, 5 ml LB and 5 µl Chloramphenicol (34 mg/ml)
09-12-16	Kim	overnightcultures	used plates from transformation (09-07-16), each plate (A3, A4, B1, B2, C2, D2) one colonie picked, 5 ml LB and 5 µl Chloramphenicol (34 mg/ml)
09-13-16	Theresa, Sabine, Wiebke	Expression	Used the overnightcultures from 09-12-16, 100 ml LB and different amounts of overnightcultures, depents on the optical density. Used 1.17 ml A, 1.8 ml B, 3.22 ml C, 2.4 ml D - Start optical density was 0.05
			Incubation; optical density after 1 h: A - 0.063, B - 0.11, C - 0.201, D - 0.115
			Optical density after 2.5 h: A - 0.452, B - 0.59, C - 0.93, D - 0.756
			Optical density after 2.75 h: A - 0.5
			Optical density after 3 h: A - 0.59
			1 ml, centrifuged, pellet stored at -20 °C (pre induction)
			add 200 mM IPTG (1 ml), incubate for 1 h and 30 min at 37 °C, 1 ml centrifuged, pellet stored at -20 °C (after induction)
			rest centrifuged 10 min, into liquid nitrogen and stored at -80 °C
09-14-16	Sabine, Wiebke, Louise, Theresa, Kim, Severin	Purification of tales	Resuspensionof sample 1.1.1 in 2ml TE-Buffer and 20 μl PMSF
			Ultrasonic treatment on ice: program9, 30%energy for 8 min (30sec on/30 sec off)
			Centrifuged at 4°C, 20.000gfor 15 min
			Supernatant transferred to column(Gravity flow Strep Tactin PepharoseColumn, Iba), purification as
			stated by iba protocol
			Flow through was stored as aliquots of 500µl labelled ‘E 1 – 6’ and ‘W 1 – 5’. Flow through of first centrifugation was stored and labelled as ‘supernatant’. Storage at -20°C.
09-15-16	Theresa, Wiebke, Kim, Sabine, Severin	SDS-Gels with samples derived from TALe 1.1.1 (09-14-2016)	50µl of each sample mixed with 5,88µl TCA (final concentration 10%). Incubated for 10 min on ice. Centrifuged at 4°C, 20 000xg for 10 min. Mixed with 500µl of 80% acetone and centrifuged at 4°C, 20 000xg for 10 min.
			Incubated for 30 min at 37°C. Mixed with 40µl Laemmli and incubated for 10 min at 80°C. Gels ran for 1 h and 20 min at 200V in TANK buffer.
		Coomassie-Staining	Coomassie-gels were set to stain over night.
		Western Blot	Other gels were Blotted semi-dry at 70V for 50 min. Poncau-staining was done for 10 min. Incubated for 1 h in PBST with 3% milkpowder. Incubation over night with 1st antibody
			(StrepMAB-Classic Cell Culture Supernatant Iga1 anti-strep-tag II monoclonal, 2-1508-025 Iba life science)
09-16-16	Sabine, Kim, Wiebke	Continuation of Western	1st antibody removed by washing in PBST. Incubation in PBST with 2nd antibody (Ecl anti-mouse IgG horseradish peroxidase) for 3 h
			Stained with 6 ml 100 mM TRIS (pH 8,5), 30 µl Luminol (22.2 mg/ 500 µl DMSO), 15 µl p-Coumaric acid (7.4 mg/ 500 µl DMSO) and 10 µl 30% H₂O₂ for 1 min up to 30 min
		Continuation of Coomassie-Staining	Destained for 1,5 h and incubated in water for 1h
09-20-16	Sabine, Wiebke, Kim, Theresa, Bianca, Louise	Purification of tales	Resuspension of sample 1.1.1, B2.1 and C2.1 in 2 ml TE-Buffer and 20 µl PMSF, Ultrasonic treatment on ice: program 9, 30% energy for 8 min (30 sec on/30 sec off). Centrifugation at 4°C, 20 000xg for 15 min
			Supernatant transferred to column (Gravity flow Strep Tactin Pepharose Colum, Iba), purification as stated by Iba protocol
			Flow through was labelled as ‘E 1 – 6’ and ‘W 1 – 5’ and stored at -20°C
		Others	Fluorecence of GFP-TAL fusion proteins in BL21 strains 1.1.1, B1 and C2 and in corresponding raw extracts was documented
		expression	Overnight cultures in 200 ml LB of BL21 strains B2 and C2 (09-12-16) inoculated and incubated at 37°C
		overnight cultures	Overnight cultures in 5ml LB and master dish of 2.1.1, 3.1.1, 3.2.1 and 4.1.1 (09-12-16) inoculated and incubated at 37°C
09-21-16	Bianca	TEV digestion	According to ProTEV plus protocol (Promega)
			2µl Buffer 0,4 µl 100mM DTT 36,8 µl Protein (E6,5) as collected on 09-14-16 0,8µl TEV
			Incubated at 30°C. Samples of 9,5µl were taken after 0, 1, 2, 4 and 6h and stored at -20°C
		Proteinexpression	Overnight cultures of BL21 strains B2 and C2 (09-20-16) were used to inoculate 5x 200ml LB+CA each and grown at 37°C to an OD600 of approx. 0,500.
			1ml samples were taken of each flask, the cells pelleted, labelled as ‘before induction’ and stored at -20°C.
			Protein expression was induced with 400 µl IPTG, final concentration: 2mM
			After 1,5h 1ml samples were taken of each flask, the cells pelleted, labelled as ‘after induction’ and stored at -20°C
			and the remaining cells were harvested and stored at -80°C
		TCA precipitation	Samples derived from 1.1.1, B2.1 and C2.1 (09-20-16) were mixed with 3 µl NaDOC and incubated for 10min on ice
			3 µl TCA were added and samples were incubated for 60 min on ice, Centrifuged at 4°C, 6 500RPM for 20 min, Pellets were resuspended in 1 ml 80% acetone,
			Centrifuged at 4 °C, 6,500RPM for 10 min, Supernatant was discarded; samples were dried at 37 °C, 30 µl SDS running-buffer + Lämmli was added, incubated at 80 °C for 10 min, Samples were stored at -20 °C
		other	Overnight culture of 1.1.1 was transferred into 30ml fresh LB+CA and incubated over night at 37 °C
09-22-16	Bianca, Janis	Proteinexpression	Overnight culture of 1.1.1 (09-21-16) was used to inoculate 5x 200ml LB+CA each and grown at 37 °C to an OD600 of approx. 0,600. 1 ml samples were taken of each flask, the cells pelleted, labelled as ‘before induction’ and stored at -20 °C.
			Proteinexpression was induced with 400 µl IPTG, final concentration: 2 mM
			After 1,5h 1 ml samples were taken of each flask, the cells pelleted, labelled as ‘after induction’ and stored at -20 °C and the remaining cells were harvested and stored at -80 °C
		SDS-gels	Gels ran for 40 min at 200V in TANK buffer
		Western Blot	semi-dry blot at 6,9 V for 45 min
			Poncau-staining was done for 10 min
			Incubation for 1 h in PBST with 3 % milkpowder
			Incubated at 4°C over night with 1st antibody (StrepMAB-Classic Cell Culture Supernatant Iga1 anti-strep-tag II monoclonal, 2-1508-025 Iba life science
			Plasmidisolation, With Quigen Mini Prep Spin Column Kit according to protocol.
		Masterdish	Petridish was damaged, redone with recent BL21 strains: A3.1, A3.2, A3.3, A4.1, A4.2, A4.3, B1.1, B1.2, B1.3, B2.1, C2.1, C2.2, C2.3, D1.1
09-23-16	Bianca, Janis, Kim	Control digest	Plasmids isolated on 09-22-16 were digested with PstI and XbaI (Set1) as well as PstI, XbaI and BsaI (Set2)
		Continuation of Western Blot	1st antibody removed by washing in PBS, Inbubation with 2nd, Incubated in PBST with 2nd antibody (EcL anti-mouse IgG horseradish peroxidase) for 2h, 2nd antibody removed by washing in PBS,
			Stained with 6 ml 100mM TRIS (pH 8,5), 30 µl Luminol (22,2 ng/500µl DMSO), 15 µl p-Coumaric acid (7,4 ng/500 µl DMSO) and 10 µl 30% H₂O₂ for 1 min up to 30 min
09-26-16	Kim, Bianca	SDS-Gel	Used premade 10% Gel from Biorad, TEV-digestion (09-21-16) samples were mixed with 10 µl Lämmli each, E6 1.1.1, B, C (09-20-16) samples were mixed with 30 µl Lämmli each, Incubation at 100°C for 10m in
			Gels ran for 45 min at 150 V in TANK buffer.
		Western Blot	Used Towbin-Buffer (25 mM Tris, 192 mM glycine, 20% (v/v) methanol (pH 8.3)) Blotted at 6V for 40min in fastBlot Trans Blot Turbo
			Poncau-staining was done for 10 min
			Incubated for 1h in blocking solution (1xPBST with 1xRotiBlock, Roth), washed with PBST
			Incubated at 4°C over night with 1st antibody
		Coomassie staining	Coomassie-gels were set to stain over night
		Overnight cultures	3.1.1 and 4.1.1 were inoculated in 25ml LB+CA and grown overnight at 37°C
09-27-16	Bianca, Kim, Wiebke	TEV digestion	According to ProTEV plus protocol (Promega)
			2µl buffer, 0,4 µl DTT, 36,8µl sample (1.1.1 E6 as collected on 09-20-16), 0,8 µl TEV
			Incubated at 30 °C. Samples of 9,5 µl were taken after 0, 1, 2, 4 and 6 h and stored at -20 °C
		continuation of Coomassie-Staining	After incubation with destaining solution no bands, not even marker, were visible. Gel discarded.
		continuation of Western Blot	1st antibody removed by washing in PBS
			Incubated in PBST with 2nd antibody (EcL anti-mouse IgG horseradish peroxidase) for 1 h
			2nd antibody removed by washing in PBS
			Stained with 6 ml 100 mM TRIS (pH 8,5), 30 µl Luminol (22,2 ng/500 µl DMSO), 15 µl p-Coumaric acid (7,4 ng/500 µl DMSO) and 10µl 30% H₂Osub>2
		Proteinexpression	Overnight cultures of BL21 3.1.1 and BL21 4.1.1 (09-26-16) was used to inoculate 50 ml LB+CA each and grown at 37 °C to an OD600 of approx. 0,600.
			1ml samples were taken of each flask, the cells pelleted, labelled as ‘before induction’ and stored at -20 °C.
			Proteinexpression was induced with 100 µl IPTG, final concentration: 2mM
			After 1,5 h 1 ml samples were taken of each flask, the cells pelleted, labelled as ‘after induction’ and stored at -20 °C and the remaining cells were harvested and stored at -80 °C
09-28-16	Kim, Bianca	Purification of tales	Resuspension of samples 1.1.1 (09-22-16), B and C (09-21-16) as well as 3.1.1 and 4.1.1 (09-27-16) in 2 ml TE-Buffer and 20 µl PMSF, Ultrasonic treatment on ice: program 9, 30% energy for 8 min (30 sec on/30 sec off)
			Centrifuged at 4 °C, 20 000xg for 15 min
			Supernatant transferred to column (Gravity flow Strep Tactin Pepharose Colum, Iba), purification as stated by Iba protocol
09-29-16	Janis, Bianca, Kim	SDS-Gel	Proben: Raw extract B from 09-21-16, E6 B from 09-20-16, E6 B from 09-28-16
			Raw extract C from 09-21-16, E6 C from 09-20-16, E6 C from 09-28-16
			Raw extract 1.1.1 from 09-22-16, E6 1.1.1 from 09-22-16, E6 1.1.1 from 09-28-16
			Raw extract 3.1.1, E6 3.1.1 from 09-28-16
			raw extract 4.1.1, E6 4.1.1 from 09-28-16
			TEV 0 from 09-27-16, TEV 1 from 09-27-16, TEV 2 from 09-27-16, TEV 4 from 09-27-16
		Coomassie-staining	Incubation overnight at RT
		Western Blot	Used Towbin-Buffer (25 mM Tris, 192 mM glycine, 20% (v/v) methanol (pH 8.3)), Blotted at 6V for 40 min in fastBlot Trans Blot Turbo
			Poncau-staining was done for 10min
			Incubated for 1h in PBST with 3% milk powder, washed with PBST
09-30-16	Kim	Continuation of Coomassie-Staining	Destained Gels
		continuation of Western Blot	Washed membrane with PBS
			Incubated with 1st antibody (StrepMAB-Classic Cell Culture Supernatant Iga1 anti-strep-tag II monoclonal, 2-1508-025 Iba life science) for 1 h
			1stAntibody removed by washing in PBS
			Inbubation with 2nd antibody (EcL anti-mouse IgG horseradish peroxidase) for one hour
			2nd Antibody removed by washing in PBS
			Stained for 1 min with 6 ml 100 mM TRIS (pH 8,5), 30 µl Luminol (22,2 ng/500 µl DMSO), 15 µl p-Coumaric acid (7,4 ng/500 µl DMSO) and 10 µl 30% H₂O₂
010-04-16	Kim, Severin, Wiebke	Colonie-PCR	Single Colonies of B, C and 1.1.1 were suspended in 10 µl H₂O and incubated at 95°C for 5min.
			1µl sample and 49µl mastermix were prepared and amplified as follows:
			50µl Buffer 5xG
			5µl dNTPs
			10µl Primer 1 (1534)
			10µl Primer 2 (1716)
			7.5µl DMSO
			135µ l H₂O
			25µl MgCl₂
			Program:
			35x (30s 98°C, 30 s 59 °C, 30 s 72 °C) and 5 min 72 °C
		agarose gel	1 % agarose Gel, Gels ran for 27min at 100V
		Proteinexpression	1ml LB+CA was inoculated with strain C as a preculture and later distributed into 10x 10ml LB.
			Cultures were grown to an OD600 of approx. 0,5 as well as 0.847
			Proteinexpression was induced at 30°C and 37°C with 2mM IPTG for as long as 3h and over night.
10-05-16	Kim, Severin, Wiebke		Liquid cultures were harvested and split into two separate reaction tubes for Dotplot and Immunostain
10-06-16	Kim, Wiebke	TEV digestion	Using samples according to ProTEV plus protocol (Promega)
			5µl Buffer
			1 µl 100 mM DTT
			40 µl Sample 1.1.1 (09-08-16) and C (09-20-16, 20 µl E5 and 20 µl E6)
			2 µl TEV
			52 µl H₂O
			Incubated at RT. Samples of 20 µl were taken after 0, 1, 2, 4 and 6h and stored at -20°C
		SDS-Gel	20µl of TEV digestion sample was mixed with 20µl Lämmli and incubated at 80°C for 10min, Gels (10%) ran for 50min at 150V in TANK buffer.
		Western Blot	Used Towbin-Buffer (25 mM Tris, 192 mM glycine, 20% (v/v) methanol (pH 8.3))
			Blotted at 25V for 40min in fastBlot Trans Blot Turbo
			Poncau-staining was done for 10min
			Washed with H₂O
			Incubated for 1h in blocking solution (1x PBST with 1x RotiBlock, Roth)
			Washed with PBS
			Incubated at 4°C over night with 1st antibody
10-07-16	Wiebke, Severin, Kim	Continue the Immunostain.	Wash the Membran with PBS
			Incubate at RT with the second antibody for one hour
			Wash with PBS
			Stained for 1min with 6 ml 100 mM TRIS (pH 8.5), 30 µl Luminol (22.2 ng/500 µl DMSO), 15 µl p-Coumaric acid (7.4 ng/500 µl DMSO) and 10 µl 30% H₂O₂
		Spotting the Oligonucleotides on a Chip	iGEM.npw as layout
			Solutions on Lafontain plates, 10 µl Oligonucleotides (100 µM) + 10 µl array it Micro spotting (2x)
			spotting
			Aftertreatment:
			1) UV-Crosslink for 3 minutes
			2) 24 hours in a dark room at room temperature (RT)
			3) reducing: 2 min 0.2 % SDS, 2x 1 min H₂O, 5 min NaBH4, 1 min icecold H₂O, 1 min 0.2 % SDS, 2x 1 min H₂O
			dry
			4) Blocking: 45 min Blocking reagent, 42 °C 5 min H₂O
			dry
		Overnightcultures	Used glycerin stock from Origami cells, incubate in 20 ml LB medium at RT over the weekend
			Used colonies from 1.1.1 and 2.1.1 from masterdish, 50 ml LB medium and 50 µl Chloramphenicol. Incubate over the weekend at RT
10-10-16	Kim, Wiebke, Severin	Purification	Samples of 1.1.1 (09-20-16) and 2.1.1 (08-30-16) were resuspended in 5ml TE-Buffer with 50 µl PMSF. Proteins were purifies according to Iba protocol
			One Column of 2.1.1 was incubated over night with 50 mM DTT. (at the 5th washing step, use 2 column bead volume (0.4 ml) and add 50 mM DTT. Incubate over night)
		Proteinexpression	Cultures of 1.1.1 and 2.1.1 were inoculated into 5x 200ml LB+CA
			Cultures were grown to an OD600 of approx. 0,5-0,6
			Proteinexpression was induced at 30°C and 37°C with 2mM IPTG for as long as 1,5h.
			Afterwards cells were harvested and stored at -80°C
		Prepation of electro competent cells	Take 1 ml or more from the overnight culture from origami cells into 50 ml LB medium. Grow the cells until they reaches a optical density at 600 nm of 0.5 – 0.6. Incubate the bacteria 20 min on ice.
			Centrifuge 10 min at 4 °C and discard the supernatant. Add 20 ml cold H₂O and resuspend the pellet. Centrifuge again 10 min at 4 °C.
			Add 20 ml cold H₂O and centrifuge for 10 min at 4 °C. Discard the supernatant and add 20 ml cold H₂O. Centrifuge for 10 min at 4 °C. Discard the supernatant and add 10 ml cold glycerin (10 %).
			Resuspend the pellet and centrifuge at 4 °C for 10 min. discard the supernatant and resuspend the pellet in 2 ml 10 % cold glycerin
			Take 100 µl aliquots into 1.5 µl reaction tubes. Froze them in liquid nitrogen and store at - 80 °C or use them directly for transformation.
		transformation	electropotation of competent origami cells with 1 µl plasmid DNA (from 1.1.1 and 2.1.1)
			incubate for 30 min in 500 µl LB at 37 °C. Plate 100 µl on LB with Chloramphenicol. incubate at 37 °C over night
10-11-16	Wiebke, Louise, Kim	Continued Purification	More samples of 1.1.1 (09-20-16) and 2.1.1 (08-30-16) were resuspended in 5 ml TE-Buffer with 50 µl PMSF. Proteins were purifies according to Iba protocol
			One Column of 1.1.1 and 2.1.1 were incubated overnight with 50 mM DTT.
			the incubate column from 10-10-16 were Eluated
		Stability test of circularised TALes	Purified protein samples of 2.1.1 were set to incubate with DTT (circularised) or without (control) for 0, 6, 24, 32, 48 h at 4 °C, RT and 37 °C.
			In addition, one set of samples was frozen at –20°C and thawed at RT for 30 min each for four continuous times.
			The linearized TAL 3HD100 was used as an additional control. After each time a fraction of 10µl was stored at -20°C for analysis.
		SDS-Gel	10µl of sample were mixed with 10µl Lämmli and incubated at 80°C for 10min.
			10µl of the sample/Lämmli mixture were put onto the Gels (10%) which ran for 50min at 150V in TANK buffer.
		western blot	Used Towbin-Buffer (25 mM Tris, 192 mM glycine, 20% (v/v) methanol (pH 8.3))
			Blotted at 25V for 40min in fastBlot Trans Blot Turbo
			Incubated for 1h in blocking solution (1x PBST with 1x RotiBlock, Roth)
			Washed with PBS
			Incubated at 4°C over night with 1st antibody
		Overnightculture	Used colonies from 1.1.1 and 2.1.1 fromtransformation with origami, 20 ml LB medium and 20 µl Chloramphenicol. Incubate at 37 °C over night
10-12-16	Wiebke, Louise, Sabine, Kim	Proteinexpression	Cultures of 1.1.1 and 2.1.1 were inoculated into 2x 125ml LB+CA
			Cultures were grown to an OD600 of approx. 0,5-0,6
			Proteinexpression was induced at 30 °C and 37 °C with 2 mM IPTG for as long as 2 h.
			Afterwards cells were harvested and stored at -80°C
		TEV digestion	Using samples 1.1.1 +/- DTT (10-11-16) and 2.1.1 +/- DTT (11-10-16) according to ProTEV plus protocol (Promega)
			Samples were taken after 0, 1, 2, 4 and 6h and stored at -20°C
		Continue Immunostain	wash membran with PBS
			Incubate second antibody for 1 hour
			wash with PBS
			Stained for 1min with 6ml 100mM TRIS (pH 8.5), 30µl Luminol (22.2ng/500µl DMSO), 15µl p-Coumaric acid (7.4ng/500µl DMSO) and 10µl 30% H₂O₂
10-13-16	Wiebke, Kim, Sabine, Louise	SDS-Gel	TEV digested samples (10-12-16) were mixed 1:1 with Lämmli and incubated at 80°C for 10min.
			Gels (10%) ran for 50min at 150V in TANK buffer.
		Stability test of circularised TALes	Purified protein samples (10-13-16), 1.1.1 (E5 + E6) and 2.1.1 (E5 + E6), were set to incubate for 0, 15 min, 5 h and 20 h at 4°C, RT and 37 °C.
			In addition, one set of samples was frozen at –20 °C and thawed at RT for 30min each for four continuous times. Each time a fraction of 10 µl was stored at -20 °C for analysis.
		Western Blot and Immunostain	Used Towbin-Buffer (25 mM Tris, 192 mM glycine, 20% (v/v) methanol (pH 8.3))
			Blotted at 25V for 40min in fastBlot Trans Blot Turbo
			Incubated for 1h in blocking solution (1x PBST with 1x RotiBlock, Roth)
			Washed with PBS
			Incubated for 3h at RT with 1st antibody (2 times Flag antibody, to times Strep antibody at TEV digest Western blot)
10-14-16	Wiebke, Kim, Sabine	Continue Immunostain	1stAntibody removed by washing in PBS
			Inbubation with 2nd antibody
			2nd Antibody removed by washing in PBS
			Stained for 1 min with 6 ml 100mM TRIS (pH 8,5), 30µl Luminol (22,2 ng/500 µl DMSO), 15 µl p-Coumaric acid (7,4 ng/500 µl DMSO) and 10 µl 30% H₂O₂
		Stability test of circularised TALes	Purified protein samples (10-13-16 and 10-14-16) of Origami with DTT were set to incubate for 6 h, 48 h ... at 4 °C, RT and 37 °C.
			In addition, one set of samples was frozen at –20 °C and thawed at RT for 30 min each for four continuous times. Each time a fraction of 10 µl was stored at -20 °C for analysis.
			Using samples E1 – E6 from 10-13-16 according to ProTEV plus protocol (Promega)
			Samples were taken after 0, 1, 2, 4 and 6h and stored at -20°C
10-16-16	Wiebke	stability Test	Probe at 4 pm (68 h for probes without DTT and 54 h for the probes with DTT)
10-17-16	Wiebke, Louise, Sabine, Theresa, Kim	SDS-gels	Used 8 SDS Gels for the probes from the two stability tests with and without DTT. One gel was used for the TEV digest.
			Gels run for 50 min at 150 V.
		Western blot	Used Towbin-Buffer (25 mM Tris, 192 mM glycine, 20% (v/v) methanol (pH 8.3))
			Blotted at 25 V for 40min in fastBlot Trans Blot Turbo
			Incubated for 1h in blocking solution (1x PBST with 1x RotiBlock, Roth)
			Washed with PBS
		Immunostain	Incubated Membrane 1-6 for 2 h at RT with 1st antibody and membrane 7-9 over night at 4 °C
			1stAntibody removed by washing in PBS
			Inbubation with 2nd antibody for 1 h
			2nd Antibody removed by washing in PBS
			Stained for 1 min with 6 ml 100mM TRIS (pH 8,5), 30 µl Luminol (22,2 ng/500 µl DMSO), 15 µl p-Coumaric acid (7,4 ng/500 µl DMSO) and 10 µl 30% H₂O₂/td>
10-18-16	Kim	Continued immunostain from 10-17-16	1stAntibody removed by washing in PBS
			Inbubation with 2nd antibody for 1 h
			2nd antibody removed by washing in PBS
			Stained for 1 min with 6 ml 100 mM TRIS (pH 8,5), 30 µl Luminol (22,2 ng/500 µl DMSO), 15 µl p-Coumaric acid (7,4 ng/500 µl DMSO) and 10 µl 30% H₂O₂

@@ Line 33: / Line 33: @@
    </tr>
    <tr>
-     <td>21.04.2016</td>
+     <td>04-21-16</td>
      <td>Bianca</td>
-     <td>prepared working area</td>
+     <td>preparation of working area</td>
-     <td>pipets and other stuff autoklaved</td>
+     <td>pipets and other stuff autoclaved</td>
    </tr>
    <tr>
-     <td>28.04.2016</td>
+     <td>04-28-16</td>
      <td>Janis, Bianca</td>
-     <td>test pipets</td>
+     <td>testing of pipets</td>
      <td>pipets work well</td>
    </tr>
@@ Line 47: / Line 47: @@
      <td></td>
      <td></td>
-     <td>Assembly of TALEs</td>
+     <td>assembly of TALEs</td>
-     <td>we cloned three different constructs in TALEs and transformed them into E.coli: 1) R-SCTALEN left HA Tag 2) R-TALEN left HA Tag 3) R-TALEN right myc Tag</td>
+     <td>we cloned three different constructs in TALEs and transformed them into <i>E.coli</i>: 1) R-SCTALEN left HA Tag 2) R-TALEN left HA Tag 3) R-TALEN right myc Tag</td>
    </tr>
    <tr>
@@ Line 54: / Line 54: @@
      <td></td>
      <td>isolation of plasmids</td>
-     <td>we isolated the two Plasmids 1) iGEM20? and 2) pET 32a+?  out of E.coli with a QIAprep(R) Spin Miniprep Kit. The concentration was 1) 354 ng/ µl and 2) 43.5 ng/ µl dsDNA measured with a NanoPhotometer</td>
+     <td>We isolated the two plasmids 1) iGEM20? and 2) pET 32a+?  out of <i>E.coli</i> with a QIAprep(R) Spin Miniprep Kit. The concentrations were 1) 354 ng/ µl and 2) 43.5 ng/ µl dsDNA measured with a NanoPhotometer</td>
    </tr>
    <tr>
-     <td>03.05.2016</td>
+     <td>05-03-16</td>
      <td>Janis</td>
-     <td>Assembly of TALEs, 2nd try</td>
+     <td>assembly of TALEs, 2nd try</td>
      <td>Cells could not be transformed, propably GGC of vectors did not work. Did the assembly again, this time over night. </td>
    </tr>
@@ Line 69: / Line 69: @@
    </tr>
    <tr>
-     <td>23.05.16</td>
+     <td>05-23-16</td>
      <td>Bianca</td>
-     <td>plasmidisolation</td>
+     <td>plasmid isolation</td>
-     <td>Plasmid isolation with QIAprep Spin Miniprep kit. The protocoll inside the kit was used without step 7. </td>
+     <td>Plasmid isolation with QIAprep Spin Miniprep kit. The protocol attached to the kit was used without step 7. </td>
    </tr>
    <tr>
@@ Line 84: / Line 84: @@
      <td></td>
      <td>control digest</td>
-     <td>control digest with plasmids (5) - (9). Mastermix for (5)-(9): 3 µl EcoRI-HF, 3 µl BamHI-HF, 12 µl buffer CutSmart and 66 µl H2O, 14 µl of the Mastermix were added to 6 µl of each plasmid DNA</td>
+     <td>Control digest with plasmids (5) - (9). Mastermix for (5)-(9): 3 µl EcoRI-HF, 3 µl BamHI-HF, 12 µl buffer CutSmart and 66 µl H<sub>2</sub>O, 14 µl of the Mastermix were added to 6 µl of each plasmid DNA</td>
    </tr>
    <tr>
@@ Line 90: / Line 90: @@
      <td></td>
      <td></td>
-     <td>control digest with plasmids (1) - (4). Mastermix for (1)-(4): 5 µl BsaI 10 µl CutSmart, 55 µl H2O, 14 µl of the Mastermix were added to 6 µl of plasmid DNA</td>
+     <td>Control digest with plasmids (1) - (4). Mastermix for (1)-(4): 5 µl BsaI 10 µl CutSmart, 55 µl H<sub>2</sub>2O, 14 µl of the Mastermix were added to 6 µl of plasmid DNA</td>
    </tr>
    <tr>
@@ Line 102: / Line 102: @@
      <td></td>
      <td>agarose gel</td>
-     <td>after the digest 5 µl loadingbuffer was add to the control digest, 20 µl were loaded on a 1 % agarose gel for 30 min and at 140 V. 7 µl of a 1 kb DNA ladder were used.</td>
+     <td>After the digest 5 µl loadingbuffer was added to the control digest, 20 µl were loaded on a 1 % agarose gel for 30 min and at 140 V. 7 µl of a 1 kb DNA ladder were used.</td>
    </tr>
    <tr>
-     <td>25.05.16</td>
+     <td>05-25-16</td>
      <td>Janis</td>
      <td>sequencing</td>
-     <td>plasmids and primer were pipet together</td>
+     <td>Plasmids and primer were pipetted.</td>
    </tr>
    <tr>
-     <td>26.05.16</td>
+     <td>05-26-16</td>
      <td>Bianca</td>
      <td>TNF reaction</td>
-     <td>500 ng DNA, 1 µl Met, ad 50 µl H2O. Used DNA: iGEM 1-2 (A and B), MPAx SCL, 90 min at 30 °C</td>
+     <td>500 ng DNA, 1 µl Met, ad 50 µl H<sub>2</sub>O. Used DNA: iGEM 1-2 (A and B), MPAx SCL, 90 min at 30 °C</td>
    </tr>
    <tr>
      <td></td>
      <td></td>
-     <td>In-vitro-cleavage assay</td>
+     <td>in-vitro-cleavage assay</td>
-     <td>TC(-): 12,5 µl DNA, 2 µl enzyme, 2 µl NEB3 buffer, 3,5 µl H2O  (+) Ax7: 10 µl DNA, 2 µl enzyme, 2 µl NEB3 buffer, 6 µl H2O; 30 min at 37 °C</td>
+     <td>TC(-): 12,5 µl DNA, 2 µl enzyme, 2 µl NEB3 buffer, 3,5 µl H<sub>2</sub>O  (+) Ax7: 10 µl DNA, 2 µl enzyme, 2 µl NEB3 buffer, 6 µl H<sub>2</sub>O; 30 min at 37 °C</td>
    </tr>
    <tr>
@@ Line 129: / Line 129: @@
    </tr>
    <tr>
-     <td>30.05.16</td>
+     <td>05-30-16</td>
      <td>Bianca</td>
-     <td>In-vitro-cleavage assay</td>
+     <td>in-vitro-cleavage assay</td>
-     <td>2 µl DNA ((+)AX7 und TC(-)), 3 µl NEB3 buffer, 5 µl BSA, 4 µl TNT(-)/Scl + 16 µl H2O or 13 µl TNT(+) + 7 µl H2O, ad 30 µl H2O</td>
+     <td>2 µl DNA ((+)AX7 und TC(-)), 3 µl NEB3 buffer, 5 µl BSA, 4 µl TNT(-)/Scl + 16 µl H<sub>2</sub>O or 13 µl TNT(+) + 7 µl HH<sub>2</sub>OO, ad 30 µl HH<sub>2</sub>OO</td>
    </tr>
    <tr>
@@ Line 138: / Line 138: @@
      <td></td>
      <td></td>
-     <td>1 h at 37 °C, 20 min at 65 °C, 10 min at 7000 xg centrifuget, 3 in at 14000 xg centrifuget, 20 µl supernatant + 5 µl LB</td>
+     <td>centrifugation for: 1 h at 37 °C, 20 min at 65 °C, 10 min at 7000xg, 3 min at 14000xg, 20 µl supernatant + 5 µl LB</td>
    </tr>
    <tr>
@@ Line 149: / Line 149: @@
      <td></td>
      <td></td>
-     <td>Western blot preparing probes</td>
+     <td>Western blot/preparing probes</td>
-     <td>at 37 °C, 30- (4 µl) and 30+ (13 µl) with 60 µl Laemmli, after 0, 30, 90 and 180 min. 10 min at 100 °C, store on ice</td>
+     <td>at 37 °C, 30- (4 µl) and 30+ (13 µl) with 60 µl Laemmli, after 0, 30, 90 and 180 min. 10 min at 100 °C, stored on ice</td>
    </tr>
    <tr>
      <td></td>
      <td></td>
-     <td>Repeat In-vitro-cleavage assay</td>
+     <td>repetition of in-vitro-cleavage assay</td>
      <td>SCL/TNT(+) with Ax7</td>
    </tr>
@@ Line 162: / Line 162: @@
      <td></td>
      <td></td>
-     <td>25 µl TNT+, 4 µl buffer, 4 µl H2O, ad 40 µl H2O</td>
+     <td>25 µl TNT+, 4 µl buffer, 4 µl H<sub>2</sub>O, ad 40 µl H<sub>2</sub>O</td>
    </tr>
    <tr>
@@ Line 168: / Line 168: @@
      <td></td>
      <td></td>
-     <td>4 µl Scl, 4 µl buffer, 25 µl H2O, ad 40 µl H2O</td>
+     <td>4 µl Scl, 4 µl buffer, 25 µl H<sub>2</sub>O, ad 40 µl H<sub>2</sub>O</td>
    </tr>
    <tr>
@@ Line 177: / Line 177: @@
    </tr>
    <tr>
-     <td>01.06.16</td>
+     <td>06-01-16</td>
      <td>Bianca</td>
      <td>SDS gel</td>
@@ Line 192: / Line 192: @@
      <td></td>
      <td></td>
-     <td>membran swing for 1 h in milkpouder and TBST, 1 x in TBST washed, 1 x 5 min in TBST washed, 1 x 10 min in TBST washed</td>
+     <td>membran swing for 1 h in milkpowder and TBST, 1 x in TBST washed, 1 x 5 min in TBST washed, 1 x 10 min in TBST washed</td>
    </tr>
    <tr>
@@ Line 198: / Line 198: @@
      <td></td>
      <td></td>
-     <td>membran with TBST and α-His (antibody) over night ( 15 µl antibody in TBST (100 µg/500 µl))</td>
+     <td>membran with TBST and α-His (antibody) over night (15 µl antibody in TBST (100 µg/500 µl))</td>
    </tr>
    <tr>
-     <td>02.06.16</td>
+     <td>06-02-16</td>
      <td>Bianca, Janis</td>
      <td></td>
@@ Line 207: / Line 207: @@
    </tr>
    <tr>
-     <td>03.06.16</td>
+     <td>06-03-16</td>
      <td>Janis</td>
      <td>transformation</td>
-     <td>chemically competent E.Coli cells were used, for Ax7L, Ax7R, scAx7L, 1-2; 2 µl DNA used and 50 µl competen cells; 20 min on ice, 45 sec at 42 °C, 1 h at 37 °C in 1 ml LB without antibitic.</td>
+     <td>chemically competent <i>E.Coli</i> cells were used, for Ax7L, Ax7R, scAx7L, 1-2; 2 µl DNA used and 50 µl competent cells; 20 min on ice, 45 sec at 42 °C, 1 h at 37 °C in 1 ml LB without antibiotic.</td>
    </tr>
    <tr>
@@ Line 216: / Line 216: @@
      <td></td>
      <td></td>
-     <td>cells with Ax7L, Ax7R, scAx7L plated on LB and ampicillin. cells with 1-2 on LB with chloramphenicol. Stored at 37 °C over night.</td>
+     <td>cells with Ax7L, Ax7R, scAx7L plated on LB and ampicillin. Cells with 1-2 on LB with chloramphenicol. Stored at 37 °C over night.</td>
    </tr>
    <tr>
-     <td>07.06.16</td>
+     <td>06-07-16</td>
      <td>Wiebke, Theresa, Sabine</td>
      <td>Interlab study</td>
-     <td>vector transformed into Top10 E.coli with the heatshock transfomation. 1 h at 37 °C </td>
+     <td>vector transformed into Top10 <i>E.coli</i> via heatshock transfomation. 1 h at 37 °C </td>
    </tr>
    <tr>
-     <td>27.06.16</td>
+     <td>06-27-16</td>
      <td>Bianca</td>
-     <td>Assembly of TALENs</td>
+     <td>assembly of TALENs</td>
      <td>new vector: p3H001; 4 rudiments with 1) Ax7L-DS, 2) Ax7R-RR, 3) Ax7L-scFOK, 4) Ax7R-scFOK</td>
    </tr>
@@ Line 234: / Line 234: @@
      <td></td>
      <td></td>
-     <td>p3H001 77,0 ng/µl, 2 µl of C-terminal and 2 µl of N-terminal module, 5 µl H2O, used PCR programm (see protocol)</td>
+     <td>p3H001 77,0 ng/µl, 2 µl of C-terminal and 2 µl of N-terminal module, 5 µl H<sub>2</sub>O, used PCR programm (see protocol)</td>
    </tr>
    <tr>
@@ Line 240: / Line 240: @@
      <td></td>
      <td>transformation</td>
-     <td>transformation into E.coli with electroporation. 200 µl LB and 30 min at 37 °C. Plated on LB chloramphenicol, stored at 37 °C over night.</td>
+     <td>transformation into <i>E.coli</i> with electroporation. 200 µl LB and 30 min at 37 °C. Plated on LB chloramphenicol, stored at 37 °C over night.</td>
    </tr>
    <tr>
-     <td>28.06.16</td>
+     <td>06-28-16</td>
      <td>Bianca</td>
      <td>overnight cultures</td>
-     <td>5 ml LB and 34 mg/ml chloramphenicol with 1 colonie from the plates. 3 culutres picked from each plate. Incubate at 37 °C over night.</td>
+     <td>5 ml LB and 34 mg/ml chloramphenicol with 1 colony from the plates. 3 cultures picked from each plate. Incubate at 37 °C over night.</td>
    </tr>
    <tr>
-     <td>30.06.16</td>
+     <td>06-30-16</td>
      <td>Bianca, Kim</td>
      <td>plasmid isolation</td>
-     <td>plasmid isolation with QIAprep Spin Miniprep kit. The protocoll inside the kit was used without step 7. Stored at -20 °C</td>
+     <td>plasmid isolation with QIAprep Spin Miniprep kit. The protocol attached to the kit was used without step 7. Stored at -20 °C</td>
    </tr>
    <tr>
@@ Line 261: / Line 261: @@
    </tr>
    <tr>
-     <td>01.07.16</td>
+     <td>07-01-16</td>
      <td>Kim</td>
      <td>control digest</td>
-     <td>control digest with the isolated plasmids from 30.06.16. 10 µl plasmid DNA, 2 µl 10x reaction buffer, 0,5 µl EcoRI, 0,5 µl PstI</td>
+     <td>control digest with the isolated plasmids from 06-30-16. 10 µl plasmid DNA, 2 µl 10x reaction buffer, 0,5 µl EcoRI, 0,5 µl PstI</td>
    </tr>
    <tr>
@@ Line 282: / Line 282: @@
      <td></td>
      <td>plasmid isolation</td>
-     <td>used the overnightcultures from 30.06.16; with QIA Prep Spin Miniprep Kit (without step 7)</td>
+     <td>used the overnightcultures from 06-30-16; with QIA Prep Spin Miniprep Kit (without step 7)</td>
    </tr>
    <tr>
-     <td>19.07.16</td>
+     <td>07-19-16</td>
      <td>Janis</td>
      <td>retransformation</td>
-     <td>plasmids 1.1, 2.1, 3.1, 3.2,  4.1 in Top10 E.coli., incubation in LB with chloramphenicol (1.1: Ax7L-DS, 2.1: Ax7R-RR, 3.1: Ax7L-scFOK, 3.2: Ax7L-scFOK, 4.1: Ax7R-scFOK)</td>
+     <td>plasmids 1.1, 2.1, 3.1, 3.2,  4.1 in Top10 <i>E.coli</i>. Incubation in LB with chloramphenicol (1.1: Ax7L-DS, 2.1: Ax7R-RR, 3.1: Ax7L-scFOK, 3.2: Ax7L-scFOK, 4.1: Ax7R-scFOK)</td>
    </tr>
    <tr>
-     <td>20.07.16</td>
+     <td>07-20-16</td>
      <td>Kim</td>
      <td>plasmid isolation</td>
-     <td>used the cultures from 19.07.16, with QIA Prep Spin Miniprep Kit (without step 7)</td>
+     <td>used the cultures from 07-19-16, with QIA Prep Spin Miniprep Kit (without step 7)</td>
    </tr>
    <tr>
-     <td>21.07.16</td>
+     <td>07-21-16</td>
      <td>Janis</td>
      <td>control digest</td>
-     <td>used plasmids from 20.06.16, with 4 µl plasmid DNA, 0,5 µl XbaI and 0,5 µl  PstI, 2 µl buffer and 10 µl H2O. For 1 h at 37 °C and 10 min at 65 °C.</td>
+     <td>used plasmids from 06-20-16, with 4 µl plasmid DNA, 0,5 µl XbaI and 0,5 µl  PstI, 2 µl buffer and 10 µl H<sub>2</sub>O. For 1 h at 37 °C and 10 min at 65 °C.</td>
    </tr>
    <tr>
-     <td>28.07.16</td>
+     <td>07-28-16</td>
      <td>Sabine, Wiebke, Theresa</td>
      <td>PCR</td>
-     <td>Run a PCR with 10 µl 5x HF Phusion Buffer, 5 µl dNTPs, 2,5 µl forward Primer, 2,5 µl reverse Primer, 1 µl template TCR5 H2, 0,5 µl Phusion polymerase, 28,5 µl H2O</td>
+     <td>run a PCR with 10 µl 5x HF Phusion Buffer, 5 µl dNTPs, 2,5 µl forward Primer, 2,5 µl reverse Primer, 1 µl template TCR5 H2, 0,5 µl Phusion polymerase, 28,5 µl H<sub>2</sub>O</td>
    </tr>
    <tr>
@@ Line 312: / Line 312: @@
      <td></td>
      <td></td>
-     <td>1. programm: 98 °C 2 min at the beginnig, denaturation at 98 °C for 10 sec, annealing at 60 °C for 10 sec, extension at 72 °C for 30 sec. steps repeated for 35 times.at the and 72 °C for 5 min. </td>
+     <td>1. programm: 98 °C 2 min at the beginnig, denaturation at 98 °C for 10 sec, annealing at 60 °C for 10 sec, extension at 72 °C for 30 sec. steps repeated for 35 times. at the and 72 °C for 5 min. </td>
    </tr>
    <tr>
@@ Line 330: / Line 330: @@
      <td></td>
      <td>transformation </td>
-     <td>1.1, 2.1, 3.1, 3.2 and 4.1 in BL21DE3 E.coli with electroporation. 1 h at 37 °C , plated on LB with chloramphenicol, inkubated at 37 °C</td>
+     <td>1.1, 2.1, 3.1, 3.2 and 4.1 in BL21DE3 <i>E.coli</i> with electroporation. 1 h at 37 °C , plated on LB with chloramphenicol, incubation at 37 °C</td>
    </tr>
    <tr>
@@ Line 336: / Line 336: @@
      <td></td>
      <td></td>
-     <td>puc57 in Top10 with heatshock at 42 °C for 40 sec,  200 µl LB added, 1 h at 37 °C. Plated on LB</td>
+     <td>puc57 in Top10 with heatshock at 42 °C for 40 sec, 200 µl LB added, 1 h at 37 °C. Plated on LB</td>
    </tr>
    <tr>
@@ Line 348: / Line 348: @@
      <td></td>
      <td></td>
-     <td>1 min at 1500 rpm, centrifuge to dry the column, 1 min at max rpm, add 15 µl EB buffer or H2O, incubate for one minute, centrifuge for 1 min at max rpm.</td>
+     <td>1 min at 1500 rpm, centrifuge to dry the column, 1 min at max rpm, add 15 µl EB buffer or H<sub>2</sub>O, incubate for one minute, centrifugation for 1 min at max rpm.</td>
    </tr>
    <tr>
      <td></td>
      <td></td>
-     <td>Cut and ligation</td>
+     <td>cut and ligation</td>
-     <td>2 µl cut smart buffer, 2 µl ATP, 2 µl puc57, 1 µl SmaI, 1 µl T4 Ligase, 15 µl Elution, 2 µl H2O, for 2 h at 30 °C</td>
+     <td>2 µl cut smart buffer, 2 µl ATP, 2 µl puc57, 1 µl SmaI, 1 µl T4 Ligase, 15 µl Elution, 2 µl H<sub>2</sub>O, for 2 h at 30 °C</td>
    </tr>
    <tr>
@@ Line 369: / Line 369: @@
    </tr>
    <tr>
-     <td>01.08.16</td>
+     <td>08-01-16</td>
      <td>Wiebke, Sabine</td>
      <td>Plasmid isolation </td>
-     <td>Plasmid isolation from overnightcultures with iGem Nterm and GFP puc57  (1, 2, 3, 4)</td>
+     <td>Plasmid isolation from overnight cultures with iGem Nterm and GFP puc57  (1, 2, 3, 4)</td>
    </tr>
    <tr>
@@ Line 384: / Line 384: @@
      <td></td>
      <td>control digest</td>
-     <td>with the isolated plasmids. used 0,5 µl BsaI, 7 µl DNA, 2 µl CutSmart buffer, ad 20 µl (10.5 µl H2O)  </td>
+     <td>with the isolated plasmids. used 0,5 µl BsaI, 7 µl DNA, 2 µl CutSmart buffer, ad 20 µl (10.5 µl H<sub>2</sub>O)  </td>
    </tr>
    <tr>
@@ Line 405: / Line 405: @@
    </tr>
    <tr>
-     <td>02.08.16</td>
+     <td>08-02-16</td>
      <td>Wiebke, Sabine, Louise</td>
      <td>plasmid isolation</td>
-     <td>Repeat the plasmid isolation from 01.08.16</td>
+     <td>Repeat the plasmid isolation from 08-01-16</td>
    </tr>
    <tr>
@@ Line 420: / Line 420: @@
      <td></td>
      <td>agarose gel</td>
-     <td>1 % agarose gel, used 7 µl 1 kb DNA ladder, 20 µl from Controll digest with 6 µl 5x LD buffer (20 µl loaded on agarose gel) </td>
+     <td>1 % agarose gel, used 7 µl 1 kb DNA ladder, 20 µl from control digest with 6 µl 5x LD buffer (20 µl loaded on agarose gel) </td>
    </tr>
    <tr>
-     <td>03.08.16</td>
+     <td>08-03-16</td>
      <td>Janis</td>
      <td></td>
@@ Line 438: / Line 438: @@
      <td></td>
      <td>transformation</td>
-     <td>puc57 with GFP - NTH3 in E.coli (Top10)</td>
+     <td>puc57 with GFP - NTH3 in <i>E.coli</i> (Top10)</td>
    </tr>
    <tr>
@@ Line 444: / Line 444: @@
      <td></td>
      <td>transformation</td>
-     <td>transformation into E.coli (BL21DE3) with electroporation. 200 µl LB and 30 min at 37 °C. Plated on LB chloramphenicol, stored at 37 °C over night.</td>
+     <td>transformation into <i>E.coli</i> (BL21DE3) with electroporation. 200 µl LB and 30 min at 37 °C. Plated on LB chloramphenicol, stored at 37 °C over night.</td>
    </tr>
    <tr>
@@ Line 453: / Line 453: @@
    </tr>
    <tr>
-     <td>10.08.16</td>
+     <td>08-10-16</td>
      <td>Kim</td>
      <td>overnightcultures</td>
-     <td>used 4 colonies from 03.08. (puc57 with GFP-NTH3 in Top10), 5 ml LB, 5 µl carbenicillin.</td>
+     <td>used 4 colonies from 08-03-16 (puc57 with GFP-NTH3 in Top10), 5 ml LB, 5 µl carbenicillin.</td>
    </tr>
    <tr>
-     <td>11.08.16</td>
+     <td>08-11-16</td>
      <td>Kim</td>
      <td>plasmid isolation</td>
-     <td>plasmid isolation with the overnightcultures from 10.08.16, used 4 ml overnightculture and QIA Prep Spin Miniprep Kit without step 7.</td>
+     <td>plasmid isolation with the overnightcultures from 08-10-16, used 4 ml overnightculture and QIA Prep Spin Miniprep Kit without step 7.</td>
    </tr>
    <tr>
-     <td>12.08.16</td>
+     <td>08-12-16</td>
      <td>Kim</td>
      <td>control digest</td>
-     <td>plasmids from 11.08.16; 5 µl DNA, 2 µl CutSmart buffer, 0.5 BsaI, 12.5 µl H2O; 1 h at 37 °C, 10 min at 65 °C</td>
+     <td>plasmids from 08-11-16; 5 µl DNA, 2 µl CutSmart buffer, 0.5 BsaI, 12.5 µl <i>E.coli</i> ; 1 h at 37 °C, 10 min at 65 °C</td>
    </tr>
    <tr>
@@ Line 477: / Line 477: @@
    </tr>
    <tr>
-     <td>16.08.16</td>
+     <td>08-16-16</td>
      <td>Theresa, Lousie, Severin</td>
      <td>PCR</td>
-     <td>like the PCR from 28.07.16</td>
+     <td>like the PCR from 07-28-16</td>
    </tr>
    <tr>
@@ Line 498: / Line 498: @@
      <td></td>
      <td>cloning</td>
-     <td>like the cloning from 03.08.16, but CurSmart buffer was used</td>
+     <td>like the cloning from 08-03-16, but CurSmart buffer was used</td>
    </tr>
    <tr>
@@ Line 504: / Line 504: @@
      <td></td>
      <td>transformation</td>
-     <td>10 µl cloning stuff, 50 µl competent E.coli cells; add 200 µl LB for 30 min</td>
+     <td>10 µl cloning stuff, 50 µl competent <i>E.coli</i> cells; add 200 µl LB for 30 min</td>
    </tr>
    <tr>
@@ Line 510: / Line 510: @@
      <td></td>
      <td></td>
-     <td>10 µl puc57, 50 µl competent E.coli cells; add 200 µl LB for 30 min</td>
+     <td>10 µl puc57, 50 µl competent <i>E.coli</i> cells; add 200 µl LB for 30 min</td>
    </tr>
    <tr>
-     <td>17.08.16</td>
+     <td>08-17-16</td>
      <td>Janis</td>
      <td>overnightcultures</td>
-     <td>5 colonies (1-5) picked from transformation (16.08.16), in 5 ml LB with 5 µl Carbenicellin</td>
+     <td>5 colonies (1-5) picked from transformation (08-16-16), in 5 ml LB with 5 µl Carbenicellin</td>
    </tr>
    <tr>
-     <td>18.08.16</td>
+     <td>08-18-16</td>
      <td>Janis</td>
      <td>plasmid isolation</td>
-     <td>plasmid isolation (overnightcultures used from 17.08.16) with QIA Prep Spin Miniprep Kit without step 7.</td>
+     <td>plasmid isolation (overnight cultures used from 08-17-16) with QIA Prep Spin Miniprep Kit without step 7.</td>
    </tr>
    <tr>
@@ Line 528: / Line 528: @@
      <td></td>
      <td>control digest</td>
-     <td>control digest with the isolated plasmids (18.06.16), same control digest like the digest from 12.08.16 (5 µl DNA, 2 µl CutSmart buffer, 0.5 BsaI, 12.5 µl H2O; 1 h at 37 °C, 10 min at 65 °C)</td>
+     <td>control digest with the isolated plasmids (06-18-16), same control digest like the digest from 08-12-16 (5 µl DNA, 2 µl CutSmart buffer, 0.5 BsaI, 12.5 µl H<sub>2</sub>O; 1 h at 37 °C, 10 min at 65 °C)</td>
    </tr>
    <tr>
@@ Line 540: / Line 540: @@
      <td></td>
      <td></td>
-     <td>1 µl enzym Mix (2 µl ATP, 2 µl NEB CutSmart, 1 µl T4-DNA Ligase, 1 µl BsaI) and 7 µl H2O</td>
+     <td>1 µl enzym Mix (2 µl ATP, 2 µl NEB CutSmart, 1 µl T4-DNA Ligase, 1 µl BsaI) and 7 µl H<sub>2</sub>O</td>
    </tr>
    <tr>
@@ Line 555: / Line 555: @@
    </tr>
    <tr>
-     <td>22.08.16</td>
+     <td>08-22-16</td>
      <td>Kim</td>
      <td>overnightcultures</td>
-     <td>picked 6 colonies from 18.08.16, with LB and Chloramphenicol</td>
+     <td>picked 6 colonies from 08-18-16, with LB and Chloramphenicol</td>
    </tr>
    <tr>
-     <td>23.08.16</td>
+     <td>08-23-16</td>
      <td>Kim</td>
      <td>plasmid isolation</td>
-     <td>plasmid isolation (used 4 ml overnightcultures from 22.08.16) with QIA Prep Spin Miniprep Kit without step 7</td>
+     <td>plasmid isolation (used 4 ml overnightcultures from 08-22-16) with QIA Prep Spin Miniprep Kit without step 7</td>
    </tr>
    <tr>
@@ Line 570: / Line 570: @@
      <td></td>
      <td>control digest</td>
-     <td>with plasmids (23.08.16), used 7 µl DNA, 2 µl 10x reaction buffer, 0.5 µl XbaI, 0.5 µl PstI, 10 µl H2O</td>
+     <td>with plasmids (08-23-16), used 7 µl DNA, 2 µl 10x reaction buffer, 0.5 µl XbaI, 0.5 µl PstI, 10 µl H<sub>2</sub>O</td>
    </tr>
    <tr>
@@ Line 585: / Line 585: @@
    </tr>
    <tr>
-     <td>24.08.16</td>
+     <td>08-24-16</td>
      <td>Janis, Kim</td>
      <td>transformation</td>
-     <td>plasmids 1.1, 2.1, 3.1, 3.2,  4.1 in BL21 E.coli., incubation on LB with Chloramphenicol at 37 °C</td>
+     <td>plasmids 1.1, 2.1, 3.1, 3.2,  4.1 in BL21 <i>E.coli</i>., incubation on LB with Chloramphenicol at 37 °C</td>
    </tr>
    <tr>
@@ Line 594: / Line 594: @@
      <td></td>
      <td>control digest</td>
-     <td>repeat control digest from 23.08.16 with 10 µl DNA (2 µl 10x reaction buffer, 0.5 µl XbaI, 0.5 µl PstI, 7 µl H2O); 1 h at 37 °C, 10 min at 65 °C</td>
+     <td>repeat control digest from 08-23-16 with 10 µl DNA (2 µl 10x reaction buffer, 0.5 µl XbaI, 0.5 µl PstI, 7 µl H<sub>2</sub>O); 1 h at 37 °C, 10 min at 65 °C</td>
    </tr>
    <tr>
@@ Line 606: / Line 606: @@
      <td></td>
      <td>overnightculures</td>
-     <td>picked 4 colonies from 18.08.16, with LB and Chloramphenicol</td>
+     <td>picked 4 colonies from 08-18-16, with LB and Chloramphenicol</td>
    </tr>
    <tr>
-     <td>25.08.16</td>
+     <td>08-25-16</td>
      <td>Janis, Kim</td>
      <td>plasmid isolation</td>
-     <td>plasmid isolation (overnightcultures used from 24.08.16 7-10(GFP-NTH3 in iGem vector)) with QIA Prep Spin Miniprep Kit without step 7.</td>
+     <td>plasmid isolation (overnightcultures used from 08-24-16 7-10(GFP-NTH3 in iGem vector)) with QIA Prep Spin Miniprep Kit without step 7.</td>
    </tr>
    <tr>
@@ Line 624: / Line 624: @@
      <td></td>
      <td>transformation</td>
-     <td>retransformation plasmids 1.1, 2.1, 3.1, 3.2 and 4.1 in Top 10 E.coli, 50 µl competent cells, 20 min on ice, 45 sec at 42 °C, 1 h at 37 °C in 200 µl LB, plated on LB with chloramphenicol</td>
+     <td>retransformation plasmids 1.1, 2.1, 3.1, 3.2 and 4.1 in Top 10 <i>E.coli</i>, 50 µl competent cells, 20 min on ice, 45 sec at 42 °C, 1 h at 37 °C in 200 µl LB, plated on LB with chloramphenicol</td>
    </tr>
    <tr>
@@ Line 630: / Line 630: @@
      <td></td>
      <td></td>
-     <td>transformation plasmids for interlab study in Top10 E.coli, 50 µl competent cells, 20 min on ice, 45 sec at 42 °C, 1h at 37 °C in 200 µl LB, plated on LB</td>
+     <td>transformation plasmids for interlab study in Top10 <i>E.coli</i>, 50 µl competent cells, 20 min on ice, 45 sec at 42 °C, 1h at 37 °C in 200 µl LB, plated on LB</td>
    </tr>
    <tr>
-     <td>26.08.16</td>
+     <td>08-26-16</td>
      <td>Kim</td>
      <td>control digest</td>
-     <td>with plasmids (25.08.16), used 11 µl DNA, 2 µl 10x reaction buffer, 0.5 µl XbaI, 0.5 µl PstI, 6 µl H2O - different XbaI and PstI stocks used, also repeat control digest with plasmids 1-6 </td>
+     <td>with plasmids (25-08-16), used 11 µl DNA, 2 µl 10x reaction buffer, 0.5 µl XbaI, 0.5 µl PstI, 6 µl H<sub>2</sub>O - different XbaI and PstI stocks used, also repeat control digest with plasmids 1-6 </td>
    </tr>
    <tr>
@@ Line 651: / Line 651: @@
    </tr>
    <tr>
-     <td>29.08.16</td>
+     <td>08-29-16</td>
      <td>Bianca</td>
      <td>Interlab study, transformation</td>
-     <td>DNA from Göttingen in 10 µl H2O; 50 µl competent cells (Top10 E.coli), 2 µl DNA, 20 min on ice, 45 sec at 42 °C, 1 ml LB 1 h at 37 °C, plated on LB with chloramphenicol, incubate at 37 °C</td>
+     <td>DNA from Göttingen in 10 µl H<sub>2</sub>O; 50 µl competent cells (Top10 <i>E.coli</i>), 2 µl DNA, 20 min on ice, 45 sec at 42 °C, 1 ml LB 1 h at 37 °C, plated on LB with chloramphenicol, incubation at 37 °C</td>
    </tr>
    <tr>
@@ Line 660: / Line 660: @@
      <td></td>
      <td>overnightcultures</td>
-     <td>3 colonies each plate (from 25.08.16, Top with 1.1, 2.1, 3.1, 4.1), with 5 ml LB and Chloramphenicol, incubate at 37 °C</td>
+     <td>3 colonies each plate (from 08-25-16, Top with 1.1, 2.1, 3.1, 4.1), with 5 ml LB and Chloramphenicol, incubate at 37 °C</td>
    </tr>
    <tr>
@@ Line 666: / Line 666: @@
      <td></td>
      <td></td>
-     <td>3 colonies each plate (from 24.08.16, BL21 with 1.1, 2.1, 3.1, 3.2, 4.1) with 5 ml LB and Chloramphenicol, incubate at 37 °C</td>
+     <td>3 colonies each plate (from 08-24-16, BL21 with 1.1, 2.1, 3.1, 3.2, 4.1) with 5 ml LB and Chloramphenicol, incubate at 37 °C</td>
    </tr>
    <tr>
-     <td>30.08.16</td>
+     <td>08-30-16</td>
      <td>Bianca, Theresa</td>
      <td>expression</td>
-     <td>2.5 ml from each overnightculture (from 29.08.16, BL21 with 1.1, 2.1, 3.1, 3.2, 4.1) in 10 ml LB</td>
+     <td>2.5 ml from each overnightculture (from 08-29-16, BL21 with 1.1, 2.1, 3.1, 3.2, 4.1) in 10 ml LB</td>
    </tr>
    <tr>
@@ Line 714: / Line 714: @@
      <td></td>
      <td>plasmid isolation</td>
-     <td>plasmid isolation (overnightcultures re-transformation from 29.08.16) with QIA Prep Spin Miniprep Kit without step 7</td>
+     <td>plasmid isolation (overnightcultures re-transformation from 08-29-16) with QIA Prep Spin Miniprep Kit without step 7</td>
    </tr>
    <tr>
@@ Line 720: / Line 720: @@
      <td></td>
      <td>overnightcultures</td>
-     <td>100 µl overnightculture (from 26.08.16 interlab study) with 8 ml LB and chloramphenicol, incubate at 37 °C</td>
+     <td>100 µl overnightculture (from 08-26-16 interlab study) with 8 ml LB and chloramphenicol, incubation at 37 °C</td>
    </tr>
    <tr>
-     <td>31.8.16</td>
+     <td>08-31-16</td>
      <td>Bianca, Theresa, Janis</td>
      <td>assembly</td>
-     <td>with 1 µl GFP-NTH3, 1 µl vector (50 ng/µl), 1 µl HexaI (Repeats 1-6), 1 µl HexaII (Repeats 7-11,5), 1 µl T4-Ligase, 1 µl BsaI, 2 µl 10mM ATP, 2 µl CutSmart buffer, ad 20 µl H2O</td>
+     <td>with 1 µl GFP-NTH3, 1 µl vector (50 ng/µl), 1 µl HexaI (Repeats 1-6), 1 µl HexaII (Repeats 7-11,5), 1 µl T4-Ligase, 1 µl BsaI, 2 µl 10mM ATP, 2 µl CutSmart buffer, ad 20 µl H<sub>2</sub>O</td>
    </tr>
    <tr>
@@ Line 789: / Line 789: @@
    </tr>
    <tr>
-     <td>01.09.16</td>
+     <td>09-01-16</td>
      <td>Janis, Theresa</td>
      <td>transformation</td>
-     <td>15 µl assambly product, 50 µl competent cells (E.coli, Top10), 20 min on ice, 45 sec at 42 °C, add 200 µl LB, 1 h at 37 °C, plated on LB with Chloramphenicol</td>
+     <td>15 µl assambly product, 50 µl competent cells (<i>E.coli</i>, Top10), 20 min on ice, 45 sec at 42 °C, add 200 µl LB, 1 h at 37 °C, plated on LB with Chloramphenicol</td>
    </tr>
    <tr>
@@ Line 804: / Line 804: @@
      <td></td>
      <td>control digest</td>
-     <td>with plasmids (25.08.16, re-transformation), used 11 µl DNA, 2 µl 10x reaction buffer, 0.5 µl XbaI, 0.5 µl PstI, 6 µl H2O</td>
+     <td>with plasmids (08-25-16re-transformation), used 11 µl DNA, 2 µl 10x reaction buffer, 0.5 µl XbaI, 0.5 µl PstI, 6 µl H<sub>2</sub>O</td>
    </tr>
    <tr>
@@ Line 813: / Line 813: @@
    </tr>
    <tr>
-     <td>05.09.16</td>
+     <td>09-05-16</td>
      <td>Theresa, Kim</td>
      <td>SDS gel</td>
@@ Line 822: / Line 822: @@
      <td></td>
      <td></td>
-     <td>separating gel (10 %): 27.25 ml H2O, 33.25 Acrylamid (30 %), 25 ml Tris (1.5 M; pH 8.8), 1 ml APS (10 %), 1 ml SDS (10 %), 40 µl TEMED</td>
+     <td>separating gel (10 %): 27.25 ml H<sub>2</sub>O, 33.25 Acrylamid (30 %), 25 ml Tris (1.5 M; pH 8.8), 1 ml APS (10 %), 1 ml SDS (10 %), 40 µl TEMED</td>
    </tr>
    <tr>
@@ Line 828: / Line 828: @@
      <td></td>
      <td></td>
-     <td>stacking gel: 34 ml H2O, 8.5 ml Acrylamid (30 %), 6.25 ml Tris (0.5 M; pH6.8), 0.5 ml APS (10 %), 0.5 ml SDS (10 %), 50 µl TEMED</td>
+     <td>stacking gel: 34 ml H<sub>2</sub>O, 8.5 ml Acrylamid (30 %), 6.25 ml Tris (0.5 M; pH 6.8), 0.5 ml APS (10 %), 0.5 ml SDS (10 %), 50 µl TEMED</td>
    </tr>
    <tr>
@@ Line 840: / Line 840: @@
      <td></td>
      <td></td>
-     <td>1. H2O, 2. Tris, 3. SDS, 4. Acrylamid, 5. APS, 6. TEMED; APS and TEMED everytime at last! After adding them be fast</td>
+     <td>1. H<sub>2</sub>O, 2. Tris, 3. SDS, 4. Acrylamid, 5. APS, 6. TEMED; APS and TEMED everytime at last! After adding them be fast</td>
    </tr>
    <tr>
@@ Line 846: / Line 846: @@
      <td></td>
      <td>overnightcultures</td>
-     <td>with colonies from tranformation 1.09.16, 4 colonies of the plates A-D, there was nothing on the plates E-H; 5 ml LB, 5 µl Chloramphenicol and the picked colonie</td>
+     <td>with colonies from tranformation 09-01-16, 4 colonies of the plates A-D, there was nothing on the plates E-H; 5 ml LB, 5 µl Chloramphenicol and the picked colonie</td>
    </tr>
    <tr>
-     <td>06.09.16</td>
+     <td>09-06-16</td>
      <td>Theresa, Kim</td>
      <td>plasmid isolation</td>
-     <td>plasmid isolation (overnightcultures  from 05.09.16, A1-3, B1-3, C1-3, D1-3) with QIA Prep Spin Miniprep Kit without step 7</td>
+     <td>plasmid isolation (overnightcultures  from 09-05-16, A1-3, B1-3, C1-3, D1-3) with QIA Prep Spin Miniprep Kit without step 7</td>
    </tr>
    <tr>
@@ Line 858: / Line 858: @@
      <td></td>
      <td>transformation</td>
-     <td>used plasmids from assembly F, G and H (31.08.16), 10 µl assambly product, 50 µl competent cells (E.coli, Top10), 20 min on ice, 45 sec at 42 °C, add 200 µl LB, 30 min at 37 °C, plated on LB with Chloramphenicol</td>
+     <td>used plasmids from assembly F, G and H (08-31-16), 10 µl assambly product, 50 µl competent cells (<i>E.coli</i>, Top10), 20 min on ice, 45 sec at 42 °C, add 200 µl LB, 30 min at 37 °C, plated on LB with Chloramphenicol</td>
    </tr>
    <tr>
-     <td>07.08.16</td>
+     <td>09-07-16</td>
      <td>Theresa, Bianca, Kim</td>
      <td>control digest</td>
-     <td>with plasmids (25.08.16), used 4 µl DNA, 2 µl 10x reaction buffer, 0.5 µl XbaI, 0.5 µl PstI, 13 µl H2O</td>
+     <td>with plasmids (08-25-16), used 4 µl DNA, 2 µl 10x reaction buffer, 0.5 µl XbaI, 0.5 µl PstI, 13 µl H<sub>2</sub>O</td>
    </tr>
    <tr>
@@ Line 882: / Line 882: @@
      <td></td>
      <td>SDS gel</td>
-     <td>prepared probes: took a little bit of the pellets (from the expression on 30.08.16), also the pre-induction and the after-induction pellets; 80 µl Leammli, resuspent, 10 min at 100 °C</td>
+     <td>prepared probes: took a little bit of the pellets (from the expression on 08-30-16), also the pre-induction and the after-induction pellets; 80 µl Leammli, resuspent, 10 min at 100 °C</td>
    </tr>
    <tr>
@@ Line 894: / Line 894: @@
      <td></td>
      <td>transformation</td>
-     <td>15 µl plasmid DNA (from plasmid isolation 06.09.16, plasmids A3, A4, B1, B2, C2, D1, D2) 50 µl competent cells (E.coli, BL21), 20 min on ice, 45 sec at 42 °C, add 200 µl LB, 35 min at 37 °C, 100 µl plated on LB with Chloramphenicol</td>
+     <td>15 µl plasmid DNA (from plasmid isolation 09-06-16, plasmids A3, A4, B1, B2, C2, D1, D2) 50 µl competent cells (<i>E.coli</i>, BL21), 20 min on ice, 45 sec at 42 °C, add 200 µl LB, 35 min at 37 °C, 100 µl plated on LB with Chloramphenicol</td>
    </tr>
    <tr>
-     <td>08.09.16</td>
+     <td>09-08-16</td>
      <td>Theresa, Kim</td>
      <td>SDS gel</td>
-     <td>gel 3 h in H2O and 4 h in destaining solution, after that over night in PBS buffer</td>
+     <td>gel 3 h in H<sub>2</sub>O and 4 h in destaining solution, after that over night in PBS buffer</td>
    </tr>
    <tr>
@@ Line 906: / Line 906: @@
      <td></td>
      <td>transformation</td>
-     <td>repeat transformation from 07.08.16, nothing changed</td>
+     <td>repeat transformation from 08-07-16, nothing changed</td>
    </tr>
    <tr>
-     <td>09.09.16</td>
+     <td>09-09-16</td>
      <td>Kim</td>
      <td>overnightcultures</td>
-     <td>used plates from transformation (07.09.16), each plate (A3, A4, B1, B2, D2) one colonie picked, 5 ml LB and 5 µl Chloramphenicol (34 mg/ml)</td>
+     <td>used plates from transformation (09-07-16), each plate (A3, A4, B1, B2, D2) one colonie picked, 5 ml LB and 5 µl Chloramphenicol (34 mg/ml)</td>
    </tr>
    <tr>
-     <td>12.09.16</td>
+     <td>09-12-16</td>
      <td>Kim</td>
      <td>overnightcultures</td>
-     <td>used plates from transformation (07.09.16), each plate (A3, A4, B1, B2, C2, D2) one colonie picked, 5 ml LB and 5 µl Chloramphenicol (34 mg/ml)</td>
+     <td>used plates from transformation (09-07-16), each plate (A3, A4, B1, B2, C2, D2) one colonie picked, 5 ml LB and 5 µl Chloramphenicol (34 mg/ml)</td>
    </tr>
    <tr>
-     <td>13.09.16</td>
+     <td>09-13-16</td>
      <td>Theresa, Sabine, Wiebke</td>
      <td>Expression</td>
-     <td>Used the overnightcultures from 12.09.16, 100 ml LB and different amounts of overnightcultures, depents on the optical density. Used 1.17 ml A, 1.8 ml B, 3.22 ml C, 2.4 ml D - Start optical density was 0.05</td>
+     <td>Used the overnightcultures from 09-12-16, 100 ml LB and different amounts of overnightcultures, depents on the optical density. Used 1.17 ml A, 1.8 ml B, 3.22 ml C, 2.4 ml D - Start optical density was 0.05</td>
    </tr>
    <tr>
@@ Line 969: / Line 969: @@
    </tr>
    <tr>
-     <td>14.09.16</td>
+     <td>09-14-16</td>
      <td>Sabine, Wiebke, Louise, Theresa, Kim, Severin</td>
      <td>Purification of tales</td>
-     <td>Resuspensionof sample 1.1.1in 2ml TE-Buffer and 20 μl PMSF</td>
+     <td>Resuspensionof sample 1.1.1 in 2ml TE-Buffer and 20 μl PMSF</td>
    </tr>
    <tr>
@@ Line 1,005: / Line 1,005: @@
    </tr>
    <tr>
-     <td>15.09.16</td>
+     <td>09-15-16</td>
      <td>Theresa, Wiebke, Kim, Sabine, Severin</td>
-     <td>SDS-Gels with samples derived from TALe 1.1.1 (14.09.2016)</td>
+     <td>SDS-Gels with samples derived from TALe 1.1.1 (09-14-2016)</td>
-     <td>50µl of each sample mixed with 5,88µl TCA (final concentration 10%). Incubated for 10 min on ice. Centrifuged at 4°C, 20.000g for 10 min. Mixed with 500µl of 80% acetone and centrifuged at 4°C, 20.000g for 10 min.</td>
+     <td>50µl of each sample mixed with 5,88µl TCA (final concentration 10%). Incubated for 10 min on ice. Centrifuged at 4°C, 20 000xg for 10 min. Mixed with 500µl of 80% acetone and centrifuged at 4°C, 20 000xg for 10 min.</td>
    </tr>
    <tr>
@@ Line 1,014: / Line 1,014: @@
      <td></td>
      <td></td>
-     <td>Incubated for 30 min at 37°C. Mixed with 40µl Laemmli and incubated for 10min at 80°C. Gels ran for 1 h and 20 min at 200V in TANK buffer. </td>
+     <td>Incubated for 30 min at 37°C. Mixed with 40µl Laemmli and incubated for 10 min at 80°C. Gels ran for 1 h and 20 min at 200V in TANK buffer. </td>
    </tr>
    <tr>
@@ Line 1,020: / Line 1,020: @@
      <td></td>
      <td>Coomassie-Staining</td>
-     <td>Coomassie-gels were set to stain over night over night.</td>
+     <td>Coomassie-gels were set to stain over night.</td>
    </tr>
    <tr>
@@ Line 1,026: / Line 1,026: @@
      <td></td>
      <td>Western Blot</td>
-     <td>Other gels were Blotted semi-dry at 70V for 50 min. Poncau-staining was done for 10min. Incubated for 1 h in PBST with 3% milkpowder, Incubated over night with 1st antibody </td>
+     <td>Other gels were Blotted semi-dry at 70V for 50 min. Poncau-staining was done for 10 min. Incubated for 1 h in PBST with 3% milkpowder. Incubation over night with 1st antibody </td>
    </tr>
    <tr>
@@ Line 1,035: / Line 1,035: @@
    </tr>
    <tr>
-     <td>16.09.16</td>
+     <td>09-16-16</td>
      <td>Sabine, Kim, Wiebke</td>
      <td>Continuation of Western</td>
-     <td>1st Antibody removed by washing in PBST, Incubated in PBST with 2nd antibody (Ecl anti-mouse IgG horseradish peroxidase) for 3 h</td>
+     <td>1st antibody removed by washing in PBST. Incubation in PBST with 2nd antibody (Ecl anti-mouse IgG horseradish peroxidase) for 3 h</td>
    </tr>
    <tr>
@@ Line 1,044: / Line 1,044: @@
      <td></td>
      <td></td>
-     <td>Stained with 6ml 100mM TRIS (pH 8,5), 30µl Luminol (22.2 mg/ 500 µl DMSO), 15µl p-Coumaric acid (7.4 mg/ 500 µl DMSO) and 10µl 30% H2O2 for 1 min up to 30 min</td>
+     <td>Stained with 6 ml 100 mM TRIS (pH 8,5), 30 µl Luminol (22.2 mg/ 500 µl DMSO), 15 µl p-Coumaric acid (7.4 mg/ 500 µl DMSO) and 10 µl 30% H<sub>2</sub>O<sub>2</sub> for 1 min up to 30 min</td>
    </tr>
    <tr>
@@ Line 1,053: / Line 1,053: @@
    </tr>
    <tr>
-     <td>20.09.16</td>
+     <td>09-20-16</td>
      <td>Sabine, Wiebke, Kim, Theresa, Bianca, Louise</td>
      <td>Purification of tales</td>
-     <td>Resuspension of sample 1.1.1, B2.1 and C2.1 in 2 ml TE-Buffer and 20 µl PMSF, Ultrasonic treatment on ice: program 9, 30% energy for 8 min (30 sec on/30 sec off), Centrifuged at 4°C, 20.000g for 15 min </td>
+     <td>Resuspension of sample 1.1.1, B2.1 and C2.1 in 2 ml TE-Buffer and 20 µl PMSF, Ultrasonic treatment on ice: program 9, 30% energy for 8 min (30 sec on/30 sec off). Centrifugation at 4°C, 20 000xg for 15 min </td>
    </tr>
    <tr>
@@ Line 1,079: / Line 1,079: @@
      <td></td>
      <td></td>
-     <td>Expression</td>
+     <td>expression</td>
-     <td>Overnight cultures in 200ml LB of BL21 strains B2 and C2 (12.09.2016) inoculated and incubated at 37°C</td>
+     <td>Overnight cultures in 200 ml LB of BL21 strains B2 and C2 (09-12-16) inoculated and incubated at 37°C</td>
    </tr>
    <tr>
      <td></td>
      <td></td>
-     <td>Overnightcultures</td>
+     <td>overnight cultures</td>
-     <td>Overnight cultures in 5ml LB and master dish of 2.1.1, 3.1.1, 3.2.1 and 4.1.1 (12.09.2016) inoculated and incubated at 37°C</td>
+     <td>Overnight cultures in 5ml LB and master dish of 2.1.1, 3.1.1, 3.2.1 and 4.1.1 (09-12-16) inoculated and incubated at 37°C</td>
    </tr>
    <tr>
-     <td>21.09.16</td>
+     <td>09-21-16</td>
      <td>Bianca</td>
      <td>TEV digestion</td>
@@ Line 1,098: / Line 1,098: @@
      <td></td>
      <td></td>
-     <td>2µl Buffer<br>0,4µl 100mM DTT<br>36,8µl Protein (E6,5) as collected on 14.09.2016<br>0,8µl TEV<br></td>
+     <td>2µl Buffer<br>0,4 µl 100mM DTT<br>36,8 µl Protein (E6,5) as collected on 09-14-16<br>0,8µl TEV<br></td>
    </tr>
    <tr>
@@ Line 1,110: / Line 1,110: @@
      <td></td>
      <td>Proteinexpression</td>
-     <td>Overnight cultures of BL21 strains B2 and C2 (20.09.2016) were used to inoculate 5x 200ml LB+CA each and grown at 37°C to an OD600 of approx. 0,500.</td>
+     <td>Overnight cultures of BL21 strains B2 and C2 (09-20-16) were used to inoculate 5x 200ml LB+CA each and grown at 37°C to an OD600 of approx. 0,500.</td>
    </tr>
    <tr>
@@ Line 1,122: / Line 1,122: @@
      <td></td>
      <td></td>
-     <td>Proteinexpression was induced with 400 µl IPTG, final concentration: 2mM</td>
+     <td>Protein expression was induced with 400 µl IPTG, final concentration: 2mM</td>
    </tr>
    <tr>
@@ Line 1,140: / Line 1,140: @@
      <td></td>
      <td>TCA precipitation</td>
-     <td>Samples derived from 1.1.1, B2.1 and C2.1 (20.09.2016) were mixed with 3µl NaDOC and incubated for 10min on ice</td>
+     <td>Samples derived from 1.1.1, B2.1 and C2.1 (09-20-16) were mixed with 3 µl NaDOC and incubated for 10min on ice</td>
    </tr>
    <tr>
@@ Line 1,146: / Line 1,146: @@
      <td></td>
      <td></td>
-     <td>3µl TCA were added and samples were incubated for 60min on ice, Centrifuged at 4°C, 6,500RPM for 20 min, Pellets were resuspended in 1ml 80% acetone, </td>
+     <td>3 µl TCA were added and samples were incubated for 60 min on ice, Centrifuged at 4°C, 6 500RPM for 20 min, Pellets were resuspended in 1 ml 80% acetone, </td>
    </tr>
    <tr>
@@ Line 1,152: / Line 1,152: @@
      <td></td>
      <td></td>
-     <td>Centrifuged at 4°C, 6,500RPM for 10 min, Supernatant was discarded; samples were dried at 37°C, 30µl SDS running-buffer + Lämmli was added, incubated at 80°C for 10min, Samples were stored at -20°C</td>
+     <td>Centrifuged at 4 °C, 6,500RPM for 10 min, Supernatant was discarded; samples were dried at 37 °C, 30 µl SDS running-buffer + Lämmli was added, incubated at 80 °C for 10 min, Samples were stored at -20 °C</td>
    </tr>
    <tr>
@@ Line 1,158: / Line 1,158: @@
      <td></td>
      <td>other</td>
-     <td>Overnight culture of 1.1.1 was transferred into 30ml fresh LB+CA and incubated over night at 37°C</td>
+     <td>Overnight culture of 1.1.1 was transferred into 30ml fresh LB+CA and incubated over night at 37 °C</td>
    </tr>
    <tr>
-     <td>22.09.16</td>
+     <td>09-22-16</td>
      <td>Bianca, Janis</td>
      <td>Proteinexpression</td>
-     <td>Overnight culture of 1.1.1 (21.09.2016) was used to inoculate 5x 200ml LB+CA each and grown at 37°C to an OD600 of approx. 0,600. 1ml samples were taken of each flask, the cells pelleted, labelled as ‘before induction’ and stored at -20°C.</td>
+     <td>Overnight culture of 1.1.1 (09-21-16) was used to inoculate 5x 200ml LB+CA each and grown at 37 °C to an OD600 of approx. 0,600. 1 ml samples were taken of each flask, the cells pelleted, labelled as ‘before induction’ and stored at -20 °C.</td>
    </tr>
    <tr>
@@ Line 1,170: / Line 1,170: @@
      <td></td>
      <td></td>
-     <td>Proteinexpression was induced with 400 µl IPTG, final concentration: 2mM</td>
+     <td>Proteinexpression was induced with 400 µl IPTG, final concentration: 2 mM</td>
    </tr>
    <tr>
@@ Line 1,176: / Line 1,176: @@
      <td></td>
      <td></td>
-     <td>After 1,5h 1ml samples were taken of each flask, the cells pelleted, labelled as ‘after induction’ and stored at -20°C and the remaining cells were harvested and stored at -80°C</td>
+     <td>After 1,5h 1 ml samples were taken of each flask, the cells pelleted, labelled as ‘after induction’ and stored at -20 °C and the remaining cells were harvested and stored at -80 °C</td>
    </tr>
    <tr>
      <td></td>
      <td></td>
-     <td>SDS-GELS</td>
+     <td>SDS-gels</td>
      <td>Gels ran for 40 min at 200V in TANK buffer</td>
    </tr>
@@ Line 1,188: / Line 1,188: @@
      <td></td>
      <td>Western Blot</td>
-     <td>semi-dry blot at 6,9V for 45 min</td>
+     <td>semi-dry blot at 6,9 V for 45 min</td>
    </tr>
    <tr>
@@ Line 1,194: / Line 1,194: @@
      <td></td>
      <td></td>
-     <td>Poncau-staining was done for 10min</td>
+     <td>Poncau-staining was done for 10 min</td>
    </tr>
    <tr>
@@ Line 1,200: / Line 1,200: @@
      <td></td>
      <td></td>
-     <td>Incubated for 1 h in PBST with 3% milkpowder</td>
+     <td>Incubation for 1 h in PBST with 3 % milkpowder</td>
    </tr>
    <tr>
@@ Line 1,221: / Line 1,221: @@
    </tr>
    <tr>
-     <td>23.09.16</td>
+     <td>09-23-16</td>
      <td>Bianca, Janis, Kim</td>
      <td>Control digest</td>
-     <td>Plasmids isolated on 22.09.2016 were digested with PstI and XbaI (Set1) as well as PstI, XbaI and BsaI (Set2)</td>
+     <td>Plasmids isolated on 09-22-16 were digested with PstI and XbaI (Set1) as well as PstI, XbaI and BsaI (Set2)</td>
    </tr>
    <tr>
@@ Line 1,230: / Line 1,230: @@
      <td></td>
      <td>Continuation of Western Blot</td>
-     <td>1stAntibody removed by washing in PBS, Inbubation with 2nd, Incubated in PBST with 2nd antibody (EcL anti-mouse IgG horseradish peroxidase) for 2h, 2nd Antibody removed by washing in PBS, </td>
+     <td>1st antibody removed by washing in PBS, Inbubation with 2nd, Incubated in PBST with 2nd antibody (EcL anti-mouse IgG horseradish peroxidase) for 2h, 2nd antibody removed by washing in PBS, </td>
    </tr>
    <tr>
@@ Line 1,236: / Line 1,236: @@
      <td></td>
      <td></td>
-     <td>Stained with 6ml 100mM TRIS (pH 8,5), 30µl Luminol (22,2ng/500µl DMSO), 15µl p-Coumaric acid (7,4ng/500µl DMSO) and 10µl 30% H2O2 for 1 min up to 30 min</td>
+     <td>Stained with 6 ml 100mM TRIS (pH 8,5), 30 µl Luminol (22,2 ng/500µl DMSO), 15 µl p-Coumaric acid (7,4 ng/500 µl DMSO) and 10 µl 30% H<sub>2</sub>O<sub>2</sub> for 1 min up to 30 min</td>
    </tr>
    <tr>
-     <td>26.09.16</td>
+     <td>09-26-16</td>
      <td>Kim, Bianca</td>
      <td>SDS-Gel</td>
-     <td>Used premade 10% Gel from Biorad, TEV-digestion (21.09.2016) samples were mixed with 10µl Lämmli each, E6 1.1.1, B, C (20.09.2016) samples were mixed with 30µl Lämmli each, Incubation at 100°C for 10min</td>
+     <td>Used premade 10% Gel from Biorad, TEV-digestion (09-21-16) samples were mixed with 10 µl Lämmli each, E6 1.1.1, B, C (09-20-16) samples were mixed with 30 µl Lämmli each, Incubation at 100°C for 10m in</td>
    </tr>
    <tr>
@@ Line 1,248: / Line 1,248: @@
      <td></td>
      <td></td>
-     <td>Gels ran for 45min at 150V in TANK buffer.</td>
+     <td>Gels ran for 45 min at 150 V in TANK buffer.</td>
    </tr>
    <tr>
@@ Line 1,260: / Line 1,260: @@
      <td></td>
      <td></td>
-     <td>Poncau-staining was done for 10min</td>
+     <td>Poncau-staining was done for 10 min</td>
    </tr>
    <tr>
@@ Line 1,287: / Line 1,287: @@
    </tr>
    <tr>
-     <td>27.09.16</td>
+     <td>09-27-16</td>
      <td>Bianca, Kim, Wiebke</td>
      <td>TEV digestion</td>
@@ Line 1,296: / Line 1,296: @@
      <td></td>
      <td></td>
-     <td>2µl Buffer, 0,4µl DTT, 36,8µl Sample (1.1.1 E6 as collected on 20.09.2016), 0,8µl TEV</td>
+     <td>2µl buffer, 0,4 µl DTT, 36,8µl sample (1.1.1 E6 as collected on 09-20-16), 0,8 µl TEV</td>
    </tr>
    <tr>
@@ Line 1,302: / Line 1,302: @@
      <td></td>
      <td></td>
-     <td>Incubated at 30°C. Samples of 9,5µl were taken after 0, 1, 2, 4 and 6h and stores at -20°C</td>
+     <td>Incubated at 30 °C. Samples of 9,5 µl were taken after 0, 1, 2, 4 and 6 h and stored at -20 °C</td>
    </tr>
    <tr>
      <td></td>
      <td></td>
-     <td>Continuation of Coomassie-Staining</td>
+     <td>continuation of Coomassie-Staining</td>
      <td>After incubation with destaining solution no bands, not even marker, were visible. Gel discarded.</td>
    </tr>
@@ Line 1,313: / Line 1,313: @@
      <td></td>
      <td></td>
-     <td>Continuation of Western Blot</td>
+     <td>continuation of Western Blot</td>
-     <td>1stAntibody removed by washing in PBS</td>
+     <td>1st antibody removed by washing in PBS</td>
    </tr>
    <tr>
@@ Line 1,320: / Line 1,320: @@
      <td></td>
      <td></td>
-     <td>Incubated in PBST with 2nd antibody (EcL anti-mouse IgG horseradish peroxidase) for 1h</td>
+     <td>Incubated in PBST with 2nd antibody (EcL anti-mouse IgG horseradish peroxidase) for 1 h</td>
    </tr>
    <tr>
@@ Line 1,326: / Line 1,326: @@
      <td></td>
      <td></td>
-     <td>2nd Antibody removed by washing in PBS</td>
+     <td>2nd antibody removed by washing in PBS</td>
    </tr>
    <tr>
@@ Line 1,332: / Line 1,332: @@
      <td></td>
      <td></td>
-     <td>Stained with 6ml 100mM TRIS (pH 8,5), 30µl Luminol (22,2ng/500µl DMSO), 15µl p-Coumaric acid (7,4ng/500µl DMSO) and 10µl 30% H2O2</td>
+     <td>Stained with 6 ml 100 mM TRIS (pH 8,5), 30 µl Luminol (22,2 ng/500 µl DMSO), 15 µl p-Coumaric acid (7,4 ng/500 µl DMSO) and 10µl 30% H<sub>2</sub>Osub>2</sub></td>
    </tr>
    <tr>
@@ Line 1,338: / Line 1,338: @@
      <td></td>
      <td>Proteinexpression</td>
-     <td>Overnight cultures of BL21 3.1.1 and BL21 4.1.1 (26.09.2016) was used to inoculate 50ml LB+CA each and grown at 37°C to an OD600 of approx. 0,600. </td>
+     <td>Overnight cultures of BL21 3.1.1 and BL21 4.1.1 (09-26-16) was used to inoculate 50 ml LB+CA each and grown at 37 °C to an OD600 of approx. 0,600. </td>
    </tr>
    <tr>
@@ Line 1,344: / Line 1,344: @@
      <td></td>
      <td></td>
-     <td>1ml samples were taken of each flask, the cells pelleted, labelled as ‘before induction’ and stored at -20°C.</td>
+     <td>1ml samples were taken of each flask, the cells pelleted, labelled as ‘before induction’ and stored at -20 °C.</td>
    </tr>
    <tr>
@@ Line 1,356: / Line 1,356: @@
      <td></td>
      <td></td>
-     <td>After 1,5h 1ml samples were taken of each flask, the cells pelleted, labelled as ‘after induction’ and stored at -20°C and the remaining cells were harvested and stored at -80°C</td>
+     <td>After 1,5 h 1 ml samples were taken of each flask, the cells pelleted, labelled as ‘after induction’ and stored at -20 °C and the remaining cells were harvested and stored at -80 °C</td>
    </tr>
    <tr>
-     <td>28.09.16</td>
+     <td>09-28-16</td>
      <td>Kim, Bianca</td>
      <td>Purification of tales</td>
-     <td>Resuspension of samples 1.1.1 (22.09.2016), B and C (21.09.2016) as well as 3.1.1 and 4.1.1 (27.09.2016) in 2 ml TE-Buffer and 20 µl PMSF, Ultrasonic treatment on ice: program 9, 30% energy for 8 min (30 sec on/30 sec off)</td>
+     <td>Resuspension of samples 1.1.1 (09-22-16), B and C (09-21-16) as well as 3.1.1 and 4.1.1 (09-27-16) in 2 ml TE-Buffer and 20 µl PMSF, Ultrasonic treatment on ice: program 9, 30% energy for 8 min (30 sec on/30 sec off)</td>
    </tr>
    <tr>
@@ Line 1,368: / Line 1,368: @@
      <td></td>
      <td></td>
-     <td>Centrifuged at 4°C, 20.000g for 15 min</td>
+     <td>Centrifuged at 4 °C, 20 000xg for 15 min</td>
    </tr>
    <tr>
@@ Line 1,377: / Line 1,377: @@
    </tr>
    <tr>
-     <td>29.09.16</td>
+     <td>09-29-16</td>
      <td>Janis, Bianca, Kim</td>
      <td>SDS-Gel</td>
-     <td>Proben: Raw extract B from 21.09., E6 B from 20.09., E6 B from 28.09.</td>
+     <td>Proben: Raw extract B from 09-21-16, E6 B from 09-20-16, E6 B from 09-28-16</td>
    </tr>
    <tr>
@@ Line 1,386: / Line 1,386: @@
      <td></td>
      <td></td>
-     <td>Raw extract C from 21.09., E6 C from 20.09., E6 C from 28.09.</td>
+     <td>Raw extract C from 09-21-16, E6 C from 09-20-16, E6 C from 09-28-16</td>
    </tr>
    <tr>
@@ Line 1,392: / Line 1,392: @@
      <td></td>
      <td></td>
-     <td>Raw extract 1.1.1 from 22.09., E6 1.1.1 from 20.09., E6 1.1.1 from 28.09.</td>
+     <td>Raw extract 1.1.1 from 09-22-16, E6 1.1.1 from 09-22-16, E6 1.1.1 from 09-28-16</td>
    </tr>
    <tr>
@@ Line 1,398: / Line 1,398: @@
      <td></td>
      <td></td>
-     <td>Raw extract 3.1.1, E6 3.1.1 from 28.09.</td>
+     <td>Raw extract 3.1.1, E6 3.1.1 from 09-28-16</td>
    </tr>
    <tr>
@@ Line 1,404: / Line 1,404: @@
      <td></td>
      <td></td>
-     <td>raw extract 4.1.1, E6 4.1.1 from 28.09</td>
+     <td>raw extract 4.1.1, E6 4.1.1 from 09-28-16</td>
    </tr>
    <tr>
@@ Line 1,410: / Line 1,410: @@
      <td></td>
      <td></td>
-     <td>TEV 0 from 27.09., TEV 1 from 27.09., TEV 2 from  27.09., TEV 4 from 27.09.</td>
+     <td>TEV 0 from 09-27-16, TEV 1 from 09-27-16, TEV 2 from  09-27-16, TEV 4 from 09-27-16</td>
    </tr>
    <tr>
      <td></td>
      <td></td>
-     <td>Coomassie-Staining</td>
+     <td>Coomassie-staining</td>
      <td>Incubation overnight at RT</td>
    </tr>
@@ Line 1,422: / Line 1,422: @@
      <td></td>
      <td>Western Blot</td>
-     <td>Used Towbin-Buffer (25 mM Tris, 192 mM glycine, 20% (v/v) methanol (pH 8.3)), Blotted at 6V for 40min in fastBlot Trans Blot Turbo</td>
+     <td>Used Towbin-Buffer (25 mM Tris, 192 mM glycine, 20% (v/v) methanol (pH 8.3)), Blotted at 6V for 40 min in fastBlot Trans Blot Turbo</td>
    </tr>
    <tr>
@@ Line 1,437: / Line 1,437: @@
    </tr>
    <tr>
-     <td>30.09.16</td>
+     <td>09-30-16</td>
      <td>Kim</td>
      <td>Continuation of Coomassie-Staining</td>
@@ Line 1,445: / Line 1,445: @@
      <td></td>
      <td></td>
-     <td>Continuation of Western Blot</td>
+     <td>continuation of Western Blot</td>
      <td>Washed membrane with PBS</td>
    </tr>
@@ Line 1,452: / Line 1,452: @@
      <td></td>
      <td></td>
-     <td>Incubated with 1st antibody (StrepMAB-Classic Cell Culture Supernatant Iga1 anti-strep-tag II monoclonal, 2-1508-025 Iba life science) for 1h</td>
+     <td>Incubated with 1st antibody (StrepMAB-Classic Cell Culture Supernatant Iga1 anti-strep-tag II monoclonal, 2-1508-025 Iba life science) for 1 h</td>
    </tr>
    <tr>
@@ Line 1,476: / Line 1,476: @@
      <td></td>
      <td></td>
-     <td>Stained for 1min with 6ml 100mM TRIS (pH 8,5), 30µl Luminol (22,2ng/500µl DMSO), 15µl p-Coumaric acid (7,4ng/500µl DMSO) and 10µl 30% H2O2</td>
+     <td>Stained for 1 min with 6 ml 100 mM TRIS (pH 8,5), 30 µl Luminol (22,2 ng/500 µl DMSO), 15 µl p-Coumaric acid (7,4 ng/500 µl DMSO) and 10 µl 30% H<sub>2</sub>O<sub>2</sub></td>
    </tr>
    <tr>
-     <td>04.10.16</td>
+     <td>010-04-16</td>
      <td>Kim, Severin, Wiebke</td>
      <td>Colonie-PCR</td>
-     <td>Single Colonies of B, C and 1.1.1 were suspended in 10µl H2O and incubated at 95°C for 5min.</td>
+     <td>Single Colonies of B, C and 1.1.1 were suspended in 10 µl H<sub>2</sub>O and incubated at 95°C for 5min.</td>
    </tr>
    <tr>
@@ Line 1,524: / Line 1,524: @@
      <td></td>
      <td></td>
-     <td>135µl H2O</td>
+     <td>135µ l H<sub>2</sub>O</td>
    </tr>
    <tr>
@@ Line 1,530: / Line 1,530: @@
      <td></td>
      <td></td>
-     <td>25µl MgCl2</td>
+     <td>25µl MgCl<sub>2</sub></td>
    </tr>
    <tr>
@@ Line 1,569: / Line 1,569: @@
    </tr>
    <tr>
-     <td>05.10.16</td>
+     <td>10-05-16</td>
      <td>Kim, Severin, Wiebke</td>
      <td></td>
@@ Line 1,575: / Line 1,575: @@
    </tr>
    <tr>
-     <td>06.10.16</td>
+     <td>10-06-16</td>
      <td>Kim, Wiebke</td>
      <td>TEV digestion</td>
@@ Line 1,590: / Line 1,590: @@
      <td></td>
      <td></td>
-     <td>1µl 100mM DTT</td>
+     <td>1 µl 100 mM DTT</td>
    </tr>
    <tr>
@@ Line 1,596: / Line 1,596: @@
      <td></td>
      <td></td>
-     <td>40µl Sample 1.1.1 (28.09.2016) and C (20.09.2016, 20µl E5 and 20µl E6)</td>
+     <td>40 µl Sample 1.1.1 (09-08-16) and C (09-20-16, 20 µl E5 and 20 µl E6)</td>
    </tr>
    <tr>
@@ Line 1,602: / Line 1,602: @@
      <td></td>
      <td></td>
-     <td>2µl TEV</td>
+     <td>2 µl TEV</td>
    </tr>
    <tr>
@@ Line 1,608: / Line 1,608: @@
      <td></td>
      <td></td>
-     <td>52µl H2O</td>
+     <td>52 µl H<sub>2</sub>O</td>
    </tr>
    <tr>
@@ Line 1,644: / Line 1,644: @@
      <td></td>
      <td></td>
-     <td>Washed with H2O</td>
+     <td>Washed with H<sub>2</sub>O</td>
    </tr>
    <tr>
@@ Line 1,665: / Line 1,665: @@
    </tr>
    <tr>
-     <td>07.10.16</td>
+     <td>10-07-16</td>
      <td>Wiebke, Severin, Kim</td>
      <td>Continue the Immunostain.</td>
@@ Line 1,674: / Line 1,674: @@
      <td></td>
      <td></td>
-     <td>Incubate at RT with the second Antibody for one hour</td>
+     <td>Incubate at RT with the second antibody for one hour</td>
    </tr>
    <tr>
@@ Line 1,686: / Line 1,686: @@
      <td></td>
      <td></td>
-     <td>Stained for 1min with 6ml 100mM TRIS (pH 8.5), 30µl Luminol (22.2ng/500µl DMSO), 15µl p-Coumaric acid (7.4ng/500µl DMSO) and 10µl 30% H2O2</td>
+     <td>Stained for 1min with 6 ml 100 mM TRIS (pH 8.5), 30 µl Luminol (22.2 ng/500 µl DMSO), 15 µl p-Coumaric acid (7.4 ng/500 µl DMSO) and 10 µl 30% H<sub>2</sub>O<sub>2</sub></td>
    </tr>
    <tr>
@@ Line 1,728: / Line 1,728: @@
      <td></td>
      <td></td>
-     <td>3) reducing: 2 min 0.2 % SDS, 2x 1 min H2O, 5 min NaBH4, 1 min icecold H2O, 1 min 0.2 % SDS, 2x 1 min H2O</td>
+     <td>3) reducing: 2 min 0.2 % SDS, 2x 1 min H<sub>2</sub>O, 5 min NaBH4, 1 min icecold H<sub>2</sub>O, 1 min 0.2 % SDS, 2x 1 min H<sub>2</sub>O</td>
    </tr>
    <tr>
@@ Line 1,740: / Line 1,740: @@
      <td></td>
      <td></td>
-     <td>4) Blocking: 45 min Blocking reagent, 42 °C 5 min H2O</td>
+     <td>4) Blocking: 45 min Blocking reagent, 42 °C 5 min H<sub>2</sub>O</td>
    </tr>
    <tr>
@@ Line 1,761: / Line 1,761: @@
    </tr>
    <tr>
-     <td>10.10.16</td>
+     <td>10-10-16</td>
      <td>Kim, Wiebke, Severin</td>
      <td>Purification</td>
-     <td>Samples of 1.1.1 (20.09.2016) and 2.1.1 (30.08.2016) were resuspended in 5ml TE-Buffer with 50µl PMSF. Proteins were purifies according to Iba protocol</td>
+     <td>Samples of 1.1.1 (09-20-16) and 2.1.1 (08-30-16) were resuspended in 5ml TE-Buffer with 50 µl PMSF. Proteins were purifies according to Iba protocol</td>
    </tr>
    <tr>
@@ Line 1,770: / Line 1,770: @@
      <td></td>
      <td></td>
-     <td>One Column of 2.1.1 was incubated over night with 50mM DTT. (at the 5th washing step, use 2 column bead volume (0.4 ml) and add 50 mM DTT. Incubate over night)</td>
+     <td>One Column of 2.1.1 was incubated over night with 50 mM DTT. (at the 5th washing step, use 2 column bead volume (0.4 ml) and add 50 mM DTT. Incubate over night)</td>
    </tr>
    <tr>
@@ Line 1,806: / Line 1,806: @@
      <td></td>
      <td></td>
-     <td>Centrifuge 10 min at 4 °C and discard the supernatant. Add 20 ml cold H2O and resuspend the pellet. Centrifuge again 10 min at 4 °C.</td>
+     <td>Centrifuge 10 min at 4 °C and discard the supernatant. Add 20 ml cold H<sub>2</sub>O and resuspend the pellet. Centrifuge again 10 min at 4 °C.</td>
    </tr>
    <tr>
@@ Line 1,812: / Line 1,812: @@
      <td></td>
      <td></td>
-     <td>Add 20 ml cold H2O and centrifuge for 10 min at 4 °C. Discard the supernatant and add 20 ml cold H2O. Centrifuge for 10 min at 4 °C. Discard the supernatant and add 10 ml cold glycerin (10 %).</td>
+     <td>Add 20 ml cold H<sub>2</sub>O and centrifuge for 10 min at 4 °C. Discard the supernatant and add 20 ml cold H<sub>2</sub>O. Centrifuge for 10 min at 4 °C. Discard the supernatant and add 10 ml cold glycerin (10 %).</td>
    </tr>
    <tr>
@@ Line 1,839: / Line 1,839: @@
    </tr>
    <tr>
-     <td>11.10.16</td>
+     <td>10-11-16</td>
      <td>Wiebke, Louise, Kim</td>
      <td>Continued Purification</td>
-     <td>More samples of 1.1.1 (20.09.2016) and 2.1.1 (30.08.2016) were resuspended in 5ml TE-Buffer with 50µl PMSF. Proteins were purifies according to Iba protocol</td>
+     <td>More samples of 1.1.1 (09-20-16) and 2.1.1 (08-30-16) were resuspended in 5 ml TE-Buffer with 50 µl PMSF. Proteins were purifies according to Iba protocol</td>
    </tr>
    <tr>
@@ Line 1,848: / Line 1,848: @@
      <td></td>
      <td></td>
-     <td>One Column of 1.1.1 and 2.1.1 were incubated over night with 50mM DTT.</td>
+     <td>One Column of 1.1.1 and 2.1.1 were incubated overnight with 50 mM DTT.</td>
    </tr>
    <tr>
@@ Line 1,854: / Line 1,854: @@
      <td></td>
      <td></td>
-     <td>the incubate column from 10.10.16 were Eluated</td>
+     <td>the incubate column from 10-10-16 were Eluated</td>
    </tr>
    <tr>
@@ Line 1,860: / Line 1,860: @@
      <td></td>
      <td>Stability test of circularised TALes</td>
-     <td>Purified protein samples of 2.1.1 were set to incubate with DTT (circularised) or without (control) for 0, 6, 24, 32, 48 h at 4°C, RT and 37°C. </td>
+     <td>Purified protein samples of 2.1.1 were set to incubate with DTT (circularised) or without (control) for 0, 6, 24, 32, 48 h at 4 °C, RT and 37 °C. </td>
    </tr>
    <tr>
@@ Line 1,866: / Line 1,866: @@
      <td></td>
      <td></td>
-     <td>In addition, one set of samples was frozen at –20°C and thawed at RT for 30min each for four continuous times.</td>
+     <td>In addition, one set of samples was frozen at –20°C and thawed at RT for 30 min each for four continuous times.</td>
    </tr>
    <tr>
@@ Line 1,923: / Line 1,923: @@
    </tr>
    <tr>
-     <td>12.10.16</td>
+     <td>10-12-16</td>
      <td>Wiebke, Louise, Sabine, Kim</td>
      <td>Proteinexpression</td>
@@ Line 1,938: / Line 1,938: @@
      <td></td>
      <td></td>
-     <td>Proteinexpression was induced at 30°C and 37°C with 2mM IPTG for as long as 2h.</td>
+     <td>Proteinexpression was induced at 30 °C and 37 °C with 2 mM IPTG for as long as 2 h.</td>
    </tr>
    <tr>
@@ Line 1,950: / Line 1,950: @@
      <td></td>
      <td>TEV digestion</td>
-     <td>Using samples 1.1.1 +/- DTT (11.10.2016) and 2.1.1 +/- DTT (11.10.2016) according to ProTEV plus protocol (Promega)</td>
+     <td>Using samples 1.1.1 +/- DTT (10-11-16) and 2.1.1 +/- DTT (11-10-16) according to ProTEV plus protocol (Promega)</td>
    </tr>
    <tr>
@@ Line 1,980: / Line 1,980: @@
      <td></td>
      <td></td>
-     <td>Stained for 1min with 6ml 100mM TRIS (pH 8.5), 30µl Luminol (22.2ng/500µl DMSO), 15µl p-Coumaric acid (7.4ng/500µl DMSO) and 10µl 30% H2O2</td>
+     <td>Stained for 1min with 6ml 100mM TRIS (pH 8.5), 30µl Luminol (22.2ng/500µl DMSO), 15µl p-Coumaric acid (7.4ng/500µl DMSO) and 10µl 30% H<sub>2</sub>O<sub>2</sub></td>
    </tr>
    <tr>
-     <td>13.10.16</td>
+     <td>10-13-16</td>
      <td>Wiebke, Kim, Sabine, Louise</td>
      <td>SDS-Gel</td>
-     <td>TEV digested samples (12.10.2016) were mixed 1:1 with Lämmli and incubated at 80°C for 10min.</td>
+     <td>TEV digested samples (10-12-16) were mixed 1:1 with Lämmli and incubated at 80°C for 10min.</td>
    </tr>
    <tr>
@@ Line 1,998: / Line 1,998: @@
      <td></td>
      <td>Stability test of circularised TALes</td>
-     <td>Purified protein samples (13.10.2016), 1.1.1 (E5 + E6) and 2.1.1 (E5 + E6), were set to incubate for  0, 15min, 5h and 20h at 4°C, RT and 37°C.</td>
+     <td>Purified protein samples (10-13-16), 1.1.1 (E5 + E6) and 2.1.1 (E5 + E6), were set to incubate for  0, 15 min, 5 h and 20 h at 4°C, RT and 37 °C.</td>
    </tr>
    <tr>
@@ Line 2,004: / Line 2,004: @@
      <td></td>
      <td></td>
-     <td>In addition, one set of samples was frozen at –20°C and thawed at RT for 30min each for four continuous times. Each time a fraction of 10µl was stored at -20°C for analysis.</td>
+     <td>In addition, one set of samples was frozen at –20 °C and thawed at RT for 30min each for four continuous times. Each time a fraction of 10 µl was stored at -20 °C for analysis.</td>
    </tr>
    <tr>
@@ Line 2,037: / Line 2,037: @@
    </tr>
    <tr>
-     <td>14.10.16</td>
+     <td>10-14-16</td>
      <td>Wiebke, Kim, Sabine</td>
      <td>Continue Immunostain</td>
@@ Line 2,058: / Line 2,058: @@
      <td></td>
      <td></td>
-     <td>Stained for 1min with 6ml 100mM TRIS (pH 8,5), 30µl Luminol (22,2ng/500µl DMSO), 15µl p-Coumaric acid (7,4ng/500µl DMSO) and 10µl 30% H2O2</td>
+     <td>Stained for 1 min with 6 ml 100mM TRIS (pH 8,5), 30µl Luminol (22,2 ng/500 µl DMSO), 15 µl p-Coumaric acid (7,4 ng/500 µl DMSO) and 10 µl 30% H<sub>2</sub>O<sub>2</sub></td>
    </tr>
    <tr>
@@ Line 2,064: / Line 2,064: @@
      <td></td>
      <td>Stability test of circularised TALes</td>
-     <td>Purified protein samples (13/14.10.2016) of Origami with DTT were set to incubate for 6 h, 48 h ... at 4°C, RT and 37°C.</td>
+     <td>Purified protein samples (10-13-16 and 10-14-16) of Origami with DTT were set to incubate for 6 h, 48 h ... at 4 °C, RT and 37 °C.</td>
    </tr>
    <tr>
@@ Line 2,070: / Line 2,070: @@
      <td></td>
      <td></td>
-     <td>In addition, one set of samples was frozen at –20°C and thawed at RT for 30min each for four continuous times. Each time a fraction of 10µl was stored at -20°C for analysis.</td>
+     <td>In addition, one set of samples was frozen at –20 °C and thawed at RT for 30 min each for four continuous times. Each time a fraction of 10 µl was stored at -20 °C for analysis.</td>
    </tr>
    <tr>
@@ Line 2,076: / Line 2,076: @@
      <td></td>
      <td></td>
-     <td>Using samples E1 – E6 from 13.10.16 according to ProTEV plus protocol (Promega)</td>
+     <td>Using samples E1 – E6 from 10-13-16 according to ProTEV plus protocol (Promega)</td>
    </tr>
    <tr>
@@ Line 2,085: / Line 2,085: @@
    </tr>
    <tr>
-     <td>16.10.16</td>
+     <td>10-16-16</td>
      <td>Wiebke</td>
-     <td>Stability Test </td>
+     <td>stability Test </td>
      <td>Probe at 4 pm (68 h for probes without DTT and 54 h for the probes with DTT)</td>
    </tr>
    <tr>
-     <td>17.10.16</td>
+     <td>10-17-16</td>
      <td>Wiebke, Louise, Sabine, Theresa, Kim</td>
-     <td>SDS gels</td>
+     <td>SDS-gels</td>
      <td>Used 8 SDS Gels for the probes from the two stability tests with and without DTT. One gel was used for the TEV digest.</td>
    </tr>
@@ Line 2,112: / Line 2,112: @@
      <td></td>
      <td></td>
-     <td>Blotted at 25V for 40min in fastBlot Trans Blot Turbo</td>
+     <td>Blotted at 25 V for 40min in fastBlot Trans Blot Turbo</td>
    </tr>
    <tr>
@@ Line 2,130: / Line 2,130: @@
      <td></td>
      <td>Immunostain </td>
-     <td>Incubated Membrane 1-6 for 2 h at RT with 1st antibody and membrane 7-9 over Night at 4 degree </td>
+     <td>Incubated Membrane 1-6 for 2 h at RT with 1st antibody and membrane 7-9 over night at 4 °C </td>
    </tr>
    <tr>
@@ Line 2,154: / Line 2,154: @@
      <td></td>
      <td></td>
-     <td>Stained for 1min with 6ml 100mM TRIS (pH 8,5), 30µl Luminol (22,2ng/500µl DMSO), 15µl p-Coumaric acid (7,4ng/500µl DMSO) and 10µl 30% H2O2</td>
+     <td>Stained for 1 min with 6 ml 100mM TRIS (pH 8,5), 30 µl Luminol (22,2 ng/500 µl DMSO), 15 µl p-Coumaric acid (7,4 ng/500 µl DMSO) and 10 µl 30% H<sub>2</sub>O<sub>2</sub>/td>
    </tr>
    <tr>
-     <td>18.10.16</td>
+     <td>10-18-16</td>
      <td>Kim</td>
-     <td>Continue Immunostain from 17.10.16</td>
+     <td>Continued immunostain from 10-17-16</td>
      <td>1stAntibody removed by washing in PBS</td>
    </tr>
@@ Line 2,172: / Line 2,172: @@
      <td></td>
      <td></td>
-     <td>2nd Antibody removed by washing in PBS</td>
+     <td>2nd antibody removed by washing in PBS</td>
    </tr>
    <tr>
@@ Line 2,178: / Line 2,178: @@
      <td></td>
      <td></td>
-     <td>Stained for 1min with 6ml 100mM TRIS (pH 8,5), 30µl Luminol (22,2ng/500µl DMSO), 15µl p-Coumaric acid (7,4ng/500µl DMSO) and 10µl 30% H2O2</td>
+     <td>Stained for 1 min with 6 ml 100 mM TRIS (pH 8,5), 30 µl Luminol (22,2 ng/500 µl DMSO), 15 µl p-Coumaric acid (7,4 ng/500 µl DMSO) and 10 µl 30% H<sub>2</sub>O<sub>2</sub></td>
    </tr>
 </table>

Difference between revisions of "Team:Hannover/Notebook"

Latest revision as of 20:55, 19 October 2016

Notebook

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Protocols