Difference between revisions of "Team:TU Darmstadt/Notebook"

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<tr><td></td><td><img class="labjournalIcon" src="https://static.igem.org/mediawiki/2016/7/7c/T--TU_Darmstadt--iconTech.png"/></td>
 
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<tr><td></td><td><img class="labjournalIcon" src="https://static.igem.org/mediawiki/2016/7/78/T--TU_Darmstadt--iconMod.png"/></td>
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<tr><td>Meeting with the people interested in the technical part of the project, discussing the task and the possible ideas.</td><td><img class="labjournalIcon" src="https://static.igem.org/mediawiki/2016/7/78/T--TU_Darmstadt--iconMod.png"/></td>
 
<td>modeling</td></tr>
 
<td>modeling</td></tr>
 
<tr><td></td><td><img class="labjournalIcon" src="https://static.igem.org/mediawiki/2016/c/ce/T--TU_Darmstadt--iconSocial.png"/></td>
 
<tr><td></td><td><img class="labjournalIcon" src="https://static.igem.org/mediawiki/2016/c/ce/T--TU_Darmstadt--iconSocial.png"/></td>

Revision as of 18:37, 19 October 2016

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iGEM TU Darmstadt 2016
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NOTEBOOK

Labjournal 2016

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
Meeting with the people interested in the technical part of the project, discussing the task and the possible ideas. 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
Lab Week 27:
October 17 - October 23

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

robotics
modeling
social