Difference between revisions of "Team:Aix-Marseille/Notebook WIP"

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<h5>Thursday, July 7th</h5>
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<ul><li>Miniprep: DesC DesB FliC coli
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Revision as of 07:55, 8 July 2016

Monday, June 13th
  • Preparation of heat competent DH5α cells
Tuesday, June 14th
  • Transformation of DH5α cells with test plasmid
Wednesday, June 15th
  • Preparation of heat competent Tg1 cells
  • Transformation of Tg1 cells with test plasmid<
Thursday, June 16th
  • Transformation of DH5α cells with pSB1C3 + RFP
  • Transformation with K880005, K823017, I0500, K1404006, B0034
Friday, June 17th
  • Miniprep K880005, K823017, I0500, K1404006, B0034
  • Miniprep pSB1C3 + RFP
Monday, June 20th
  • pSB1C3 digestion from 17/06 miniprep
  • pSB1C3 digestion gel migration : NO PHOTO
  • Biobricks minipreps gel migration : OK PHOTO
  • Transformation Tg1 with RFP+pSB1C3
Tuesday, June 21th
  • pSB1C3 digestion and gel migration, NucleoSpin gel extract and PCR cleanup
  • FliC E.Coli and FliC Desulfo digestion E/P and ligation with pSB1C3
  • E. Coli and Desulfo FliC ligation in linearized pSB1C3
  • Transformation of pSB1C3 RFP in Tg1
Wednesday, June 22th
  • Transformation in DH5α of :
    • FliC coli/desulfo in pSB1C3
    • Linearized pSB1K3
    • Linearized pSB1A3
    • Linearized pSB1T3
    • Linearized pSB1C3
  • Miniprep B0034
  • Transformation in DH5α of interlab studies plasmids
Thursday, June 23th
  • Digestion E/P pSB1C3
  • Purification pSB1C3 purified linearized 23/06
  • Digestion E/P of DesA, DesB, DesC, DesD
  • 3A Assembly of B0034 and I0500 in pSB1K3
  • Digestion with pSB1C3 23/06 insert digested fliC
  • Transformation in Tg1 and DH5α
Friday, June 24th
  • Digestion E/P pSB1C3, pSB1A3, pSB1K3, pSB1T3, and agarose gel extract
  • Colony PCR of
    • RBS+promoter
    • GFP
    • empty
  • Ligation: FliC E.Coli, FliC desulfo, DesA B C D
  • Ligation pSB1C3 and fliC E.Coli / desulfo digested E/P 21/06
  • Transformation in Tg1 and DH5α
  • Redone transformation test device 1 and 3 interlab studies
  • Miniprep pSB1C3 A3 K3 T3
Monday, June 27th
  • Transformations DesA, DesB, DesC, DesD, FliC E.Coli and desulfo, interlab studies
  • Transformations failed
  • Transformation pSB1T3 miniprep, pSB1K3 miniprep, control
Tuesday, June 28th
  • Miniprep promoter+RBS
  • PCR of IDT sequences (fliC and DesA to D) and gel check
  • Gel extract K3, C3, T3
Wednesday, June 29th
  • Miniprep from interlab studies, T3 and K3
  • Ligation and transformation in Tg1 Sandra Des A to D, fliC coli desulfo
  • Redone PCR of IDT sequences (fliC and DesA to D) (different Tm) and gel extract
  • Transformation of fliCs from remaining ligation in Tg1 Sandra
Thursday, June 30th
  • Migration FliC coli, desulfo, DesB, DesC
Friday, July 1st
  • Digestion PCR products and vector pSB1C3
  • PCR clean up on digested pSB1C3, fliC coli desulfo, DesB and DesC, migration and transformation in Tg1
Monday, July 4th
  • Isolated white colonies on dishes
Tuesday, July 5th
  • Reisolated colonies from 04/07
Wednesday, July 6th
  • Colony PCR with FLiC E.Coli Desulfo DesB DesC
Thursday, July 7th
  • Miniprep: DesC DesB FliC coli