Difference between revisions of "Team:Aix-Marseille/Notebook WIP"
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<h5>Tuesday, June 21th</h5> | <h5>Tuesday, June 21th</h5> | ||
<ul> | <ul> | ||
| − | <li>pSB1C3 digestion and gel migration, NucleoSpin gel | + | <li>pSB1C3 digestion and gel migration, NucleoSpin gel extract and PCR cleanup</li> |
<li>FliC E.Coli and FliC Desulfo digestion E/P and ligation with pSB1C3</li> | <li>FliC E.Coli and FliC Desulfo digestion E/P and ligation with pSB1C3</li> | ||
<li>E. Coli and Desulfo FliC ligation in linearized pSB1C3</li> | <li>E. Coli and Desulfo FliC ligation in linearized pSB1C3</li> | ||
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<h5>Friday, June 24th</h5> | <h5>Friday, June 24th</h5> | ||
| − | <ul><li>Digestion E/P pSB1C3, pSB1A3, pSB1K3, pSB1T3, and agarose gel | + | <ul><li>Digestion E/P pSB1C3, pSB1A3, pSB1K3, pSB1T3, and agarose gel extract</li> |
<li>Colony PCR of <ul><li>RBS+promoter<li>GFP<li>empty</ul></li> | <li>Colony PCR of <ul><li>RBS+promoter<li>GFP<li>empty</ul></li> | ||
<li>Ligation: FliC E.Coli, FliC desulfo, DesA B C D</li> | <li>Ligation: FliC E.Coli, FliC desulfo, DesA B C D</li> | ||
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<h5>Tuesday, June 28th</h5> | <h5>Tuesday, June 28th</h5> | ||
<ul><li>Miniprep promoter+RBS</li> | <ul><li>Miniprep promoter+RBS</li> | ||
| − | <li>PCR of IDT sequences (fliC and DesA to D) | + | <li>PCR of IDT sequences (fliC and DesA to D) and gel check |
| − | <li>Gel | + | <li>Gel extract K3, C3, T3</li> |
</ul> | </ul> | ||
<h5>Wednesday, June 29th</h5> | <h5>Wednesday, June 29th</h5> | ||
<ul><li>Miniprep from interlab studies, T3 and K3 | <ul><li>Miniprep from interlab studies, T3 and K3 | ||
| − | <li>Ligation and transformation Des A to D, fliC coli desulfo | + | <li>Ligation and transformation in Tg1 Sandra Des A to D, fliC coli desulfo |
| − | <li> | + | <li>Redone PCR of IDT sequences (fliC and DesA to D) (different Tm) and gel extract |
| − | <li> | + | <li>Transformation of fliCs from remaining ligation in Tg1 Sandra |
</ul> | </ul> | ||
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<li>PCR clean up on digested pSB1C3, fliC coli desulfo, DesB and DesC, migration and transformation in Tg1 | <li>PCR clean up on digested pSB1C3, fliC coli desulfo, DesB and DesC, migration and transformation in Tg1 | ||
</ul> | </ul> | ||
| − | |||
<h5>Monday, July 4th</h5> | <h5>Monday, July 4th</h5> | ||
Revision as of 14:47, 6 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/li>
- 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
