Difference between revisions of "Team:Aix-Marseille/Notebook WIP"
| Line 76: | Line 76: | ||
<li>Transformation in Tg1 and DH5α | <li>Transformation in Tg1 and DH5α | ||
<li>Redone transformation test device 1 and 3 interlab studies | <li>Redone transformation test device 1 and 3 interlab studies | ||
| − | <li>Miniprep pSB1C3 | + | <li>Miniprep pSB1C3 pSB1CA3 pSB1CK3 pSB1CT3 |
</ul> | </ul> | ||
| Line 88: | Line 88: | ||
<ul><li>Miniprep promoter+RBS | <ul><li>Miniprep promoter+RBS | ||
<li>PCR of IDT sequences (fliC and DesA to D) and gel check | <li>PCR of IDT sequences (fliC and DesA to D) and gel check | ||
| − | <li>Gel extract | + | <li>Gel extract pSB1CK3, pSB1CC3, pSB1CT3 |
</ul> | </ul> | ||
| Line 122: | Line 122: | ||
<h5>Thursday, July 7th</h5> | <h5>Thursday, July 7th</h5> | ||
<ul><li>Miniprep: DesC DesB FliC coli | <ul><li>Miniprep: DesC DesB FliC coli | ||
| + | <li>PCR on A2 A4 B1 clones from 6/7 | ||
| + | <li>Digestion E/P of A5 B4 C3 C2 minipreps and pSB1A3, pSB1C3, pSB1K3, pSB1T3 | ||
| + | </ul> | ||
| + | |||
| + | <h5>Friday, July 8th</h5> | ||
| + | <ul><li>Transformation BOO34 and I0500 in pSB1K3 | ||
| + | <li>Gel verification of 7/7 digestion | ||
| + | <li>Miniprep A5 (fliC coli) and C2 (DesB) from 7/7 starters | ||
| + | <li>PCR clean up of vectors | ||
| + | <li>PCR SLIC FliC coli/desulfo, DesA, B, C, D, csgA -> failed | ||
| + | </ul> | ||
| + | |||
| + | <h5>Monday, July 11th</h5> | ||
| + | <ul><li>Redone PCR SLIC 8/8 -> ok | ||
| + | <li>Redone PCR DesC, DesD and csgA | ||
| + | <li>SLIC desA, desB, fliC coli desulfo in pSB1A3 | ||
| + | <li>Transformation Tg1 with fliC coli desulfo, desA, desB in pSB1A3 from SLIC | ||
| + | <li>PCR clean up desA, desB, fliC coli desulfo | ||
</ul> | </ul> | ||
</div> | </div> | ||
</html> | </html> | ||
Latest revision as of 08:27, 12 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 pSB1CA3 pSB1CK3 pSB1CT3
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 pSB1CK3, pSB1CC3, pSB1CT3
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
- PCR on A2 A4 B1 clones from 6/7
- Digestion E/P of A5 B4 C3 C2 minipreps and pSB1A3, pSB1C3, pSB1K3, pSB1T3
Friday, July 8th
- Transformation BOO34 and I0500 in pSB1K3
- Gel verification of 7/7 digestion
- Miniprep A5 (fliC coli) and C2 (DesB) from 7/7 starters
- PCR clean up of vectors
- PCR SLIC FliC coli/desulfo, DesA, B, C, D, csgA -> failed
Monday, July 11th
- Redone PCR SLIC 8/8 -> ok
- Redone PCR DesC, DesD and csgA
- SLIC desA, desB, fliC coli desulfo in pSB1A3
- Transformation Tg1 with fliC coli desulfo, desA, desB in pSB1A3 from SLIC
- PCR clean up desA, desB, fliC coli desulfo
