Team:TU Darmstadt/Notebook

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NOTEBOOK

See Methodic Protocols and Materials →

Labjournal 2016

Lab Weeks

1 (04/18 - 04/24)	2 (04/25 - 05/01)	3 (05/02 - 05/08)	4 (05/09 - 05/15)	5 (05/16 - 05/22)
6 (05/23 - 05/29)	7 (05/30 - 06/05)	8 (06/06 - 06/12)	9 (06/13 - 06/19)	10 (06/20 - 06/26)
11 (06/27 - 07/03)	12 (07/04 - 07/10)	13 (07/11 - 07/17)	14 (07/18 - 07/24)	15 (07/25 - 07/31)
16 (08/01 - 08/07)	17 (08/08 - 08/14)	18 (08/15 - 08/21)	19 (08/22 - 08/28)	20 (08/29 - 09/04)
21 (09/05 - 09/11)	22 (09/12 - 09/18)	23 (09/19 - 09/25)	24 (09/26 - 10/02)	25 (10/03 - 10/09)
26 (10/10 - 10/16)	27 (10/17 - 10/23)

For half a year we were working in the wet lab    , developed a pipetting robot   and simulated the molecular dynamics of our proteins    . As well we would like to highlight certain social events   which helped us to further improve our team spirit.

Lab Week 1: April 18 - April 24		Hier könnte Ihr labjournal stehen!
		robotics
		modeling
		social

Lab Week 2: April 25 - Mai 1		Hier könnte Ihr labjournal stehen!
		robotics
		modeling
		social

Lab Week 3: May 2 - May 8		Hier könnte Ihr labjournal stehen!
		robotics
		modeling
		social

Lab Week 4: May 9 - May 15		Hier könnte Ihr labjournal stehen!
		robotics
		modeling
		social

Lab Week 5: May 16 - May 22		Hier könnte Ihr labjournal stehen!
		robotics
		modeling
		social

Lab Week 6: May 23 - May 29		Hier könnte Ihr labjournal stehen!
		robotics
		modeling
		social

Lab Week 7: May 30 - June 5		Hier könnte Ihr labjournal stehen!
		robotics
		modeling
		social

Lab Week 8: June 6 - June 12		Hier könnte Ihr labjournal stehen!
		robotics
		modeling
		social

Lab Week 9: June 13 - June 19		Hier könnte Ihr labjournal stehen!
		robotics
		modeling
		social

Lab Week 10: June 20 - June 26		Hier könnte Ihr labjournal stehen!
		robotics
		modeling
		social

Lab Week 11: June 27 - July 3		Hier könnte Ihr labjournal stehen!
		robotics
		modeling
		social

Lab Week 12: July 4 - July 10		Hier könnte Ihr labjournal stehen!
		robotics
		modeling
		social

Lab Week 13: July 11 - July 17		Hier könnte Ihr labjournal stehen!
		robotics
		modeling
		social

Lab Week 14: July 18 - July 24		Hier könnte Ihr labjournal stehen!
		robotics
		modeling
		social

Lab Week 15: July 25 - July 31		Hier könnte Ihr labjournal stehen!
		robotics
		modeling
		social

Lab Week 16: August 1 - August 7		Hier könnte Ihr labjournal stehen!
		robotics
		modeling
		social

Lab Week 17: August 8 - August 14		Reporter System BBa_K1976002: The mVenus-LVA-gene was synthesized by idt and amplified through PCR using the SP-amplify-fw-Rep1 (fw) and SP-amplify-rev-Rep2 (REV) oligonucleotides.
		robotics
		modeling
		social

Lab Week 18: August 15 - August 21		Reporter System BBa_K1976002: The PCR product was verified by agarose gel electrophoresis and subsequently purified. The mVenus amplicon was cut with EcoRI and PstI and ligated into a pSB1C3 vector using T4-ligase.
		robotics
		modeling
		social

Lab Week 19: August 22 - August 28		Reporter System BBa_K1976002: The ligation product was transformed into E.coli Top 10 by heat shock transformation and the cells were spread out on a CMP agar plate.
		robotics
		modeling
		social

Lab Week 20: August 29 - September 4		Reporter System BBa_K1976002: Clones were screened with colony PCR using VF2 and VR oligonucleotides. Plasmid DNA was isolated from positive clones. Metabolic burden BBa_K1976001: The BioBrick BBa_I11020 was synthezised with a LVA‑tag and with B0034 as a ribosome binding site by IDT.
		robotics
		modeling
		social

Lab Week 21: September 5 - September 11		Reporter System BBa_K1976002: Isolated Plasmid DNA Sequence was verified by Sanger Sequencing. Metabolic burden BBa_K1976001: The LVA‑tag was deleted by PCR with the primer Del_LVA_fw and Del_LVA_rev.
		robotics
		modeling
		social

Lab Week 22: September 12 - September 18		Reporter System BBa_K1976003: The mVenus-LVA-pSB1C3 plasmid was cut with EcoRI and PstI and ligated into a T7-RBS-pSB1A3 plasmid which was cut with SpeI and PstI. Metabolic burden BBa_K1976000: The BioBrick BBa_E0240 was cut with the restriction enzymes XbaI and PstI. BBa_J61002 was cut with SpeI and PstI. The two restriction products were ligated by standard T4‑Ligation protocol. BBa_K1976001: B0034‑BBa_I11020 was cut with XbaI and PstI. BBa_J23101 was cut with SpeI and PstI. Both fragments were ligated using the T4‑Ligase by standard protocol.
		robotics
		modeling
		social

Lab Week 23: September 19 - September 25		Reporter System BBa_K1976003: The ligation product was transformed into E.coli Top 10 by heat shock transformation and the cells were spread out on a AMP agar plate. Metabolic burden BBa_K1976000: After plasmid preparation, the plasmid was cut with XbaI and PstI. We had the attp‑site BBa_11023 flanked by the two additional bidirectional transcription terminators BBa_1001 synthesized by IDT. The attp‑site of BBa_11023 was not the original λ‑attp‑site. Instead the sequence corresponded to the attP2‑site used in the Gateway ® cloning system. The construct was cut with EcoRI and SpeI. The pSB1C3 backbone was cut with EcoRI and PstI. Together with the previously cut BBa_J61002‑BBa_E0240 construct the synthesized BBa_1001‑BBa_11023‑BBa_1001 construct was ligated in the cut pSB1C3. BBa_K1976001: The ligation product was transformed into E. coli Top 10 via heatshock.
		robotics
		modeling
		social

Lab Week 24: September 26 - October 2		Reporter System BBa_K1976003: Clones were screened with colony PCR using VF2 and VR oligonucleotides. Plasmid DNA was isolated from positive clones . Metabolic burden BBa_K1976000: The attP2‑site was mutated to λ‑attp via Quick‑Change‑PCR, using the following primers: QC_attp_fw and QC_attp_rev (for more details see our primerlist). BBa_K1976001: To check whether BBa_I11020 was expressed properly a SDS‑Page was made.
		robotics
		modeling
		social

Lab Week 25: October 3 - October 9		Reporter System BBa_K1976003: The isolated Plasmid DNA was verified by Sanger sequencing. Metabolic burden BBa_K1976000: The LVA‑tag was added by PCR with the overhang primer LVA_GFP_fw and LVA_GFP_rev.
		robotics
		modeling
		social

Lab Week 26: October 10 - October 16		Reporter System BBa_K1976003: Subsequently the psB1A3 vector containing brick was cut with EcoRI and PstI and ligated into a pSB1C3 vector using T4‑Ligase. Metabolic burden BBa_K1976000: For plasmid curing the final plasmid construct and the low copy backbone pSB4A5 were cut with EcoRI and PstI and then ligated. BBa_K1976001: For plasmid curing the final plasmid construct and the low copy vector pSB4K5 were cut with EcoRI and PstI and then ligated. BBa_K1976000 and BBa_K1976001 were cotransformed into E. coli Top 10. To check whether the genomic integration was successful a cPCR with attb_fw and VR primers was made.
		robotics
		modeling
		social

Metabolic burden

Plasmid curing could not be performed due to lack of time.

Lab Week 27: October 17 - October 23
		robotics
		modeling
		social

iGEM Technische Universität Darmstadt

We thank our sponsors, especially:

Email: igem@bio.tu-darmstadt.de | Design: Viktoria Schuster