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

 
(13 intermediate revisions by the same user not shown)
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<h5>Tuesday, June 14th</h5>
 
<h5>Tuesday, June 14th</h5>
 
<ul>
 
<ul>
−
<li>Transformation of DH5α cells with test plasmid</li>
+
<li>Transformation of DH5α cells with test plasmid
 
</ul>
 
</ul>
  
 
<h5>Wednesday, June 15th</h5>
 
<h5>Wednesday, June 15th</h5>
 
<ul>
 
<ul>
−
<li>Preparation of heat competent Tg1 cells</li>
+
<li>Preparation of heat competent Tg1 cells
−
<li>Transformation of Tg1 cells with test plasmid</li>
+
<li>Transformation of Tg1 cells with test plasmid<
 
</ul>
 
</ul>
  
 
<h5>Thursday, June 16th</h5>
 
<h5>Thursday, June 16th</h5>
 
<ul>
 
<ul>
−
<li>Transformation of DH5α cells with pSB1C3 + RFP/li>
+
<li>Transformation of DH5α cells with pSB1C3 + RFP
−
<li>Transformation with K880005, K823017, I0500, K1404006, B0034</li>
+
<li>Transformation with K880005, K823017, I0500, K1404006, B0034
 
</ul>
 
</ul>
 
<h5>Friday, June 17th</h5>
 
<h5>Friday, June 17th</h5>
 
<ul>
 
<ul>
−
<li>Miniprep K880005, K823017, I0500, K1404006, B0034</li>
+
<li>Miniprep K880005, K823017, I0500, K1404006, B0034
−
<li>Miniprep pSB1C3 + RFP</li>
+
<li>Miniprep pSB1C3 + RFP
 
</ul>
 
</ul>
  
 
<h5>Monday, June 20th</h5>
 
<h5>Monday, June 20th</h5>
 
<ul>
 
<ul>
−
<li>pSB1C3 digestion from 17/06 miniprep</li>
+
<li>pSB1C3 digestion from 17/06 miniprep
−
<li>pSB1C3 digestion gel migration : NO PHOTO</li>
+
<li>pSB1C3 digestion gel migration : NO PHOTO
−
<li>Biobricks minipreps gel migration : OK PHOTO</li>
+
<li>Biobricks minipreps gel migration : OK PHOTO
−
<li>Transformation Tg1 with RFP+pSB1C3</li>
+
<li>Transformation Tg1 with RFP+pSB1C3
 
</ul>
 
</ul>
  
 
<h5>Tuesday, June 21th</h5>
 
<h5>Tuesday, June 21th</h5>
 
<ul>
 
<ul>
−
<li>pSB1C3 digestion and gel migration, NucleoSpin gel purification and PCR cleanup</li>
+
<li>pSB1C3 digestion and gel migration, NucleoSpin gel extract and PCR cleanup
−
<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>E. Coli and Desulfo FliC ligation in linearized pSB1C3</li>
+
<li>E. Coli and Desulfo FliC ligation in linearized pSB1C3
−
<li>Transformation of pSB1C3 RFP in Tg1</li>
+
<li>Transformation of pSB1C3 RFP in Tg1
−
 
+
</ul>
 
<h5>Wednesday, June 22th</h5>
 
<h5>Wednesday, June 22th</h5>
 
<ul>
 
<ul>
 
<li>Transformation in DH5α of :<ul>
 
<li>Transformation in DH5α of :<ul>
−
<li>FliC coli/desulfo in pSB1C3</li>
+
<li>FliC coli/desulfo in pSB1C3
−
<li>Linearized pSB1K3</li>
+
<li>Linearized pSB1K3
−
<li>Linearized pSB1A3</li>
+
<li>Linearized pSB1A3
−
<li>Linearized pSB1T3</li>
+
<li>Linearized pSB1T3
−
<li>Linearized pSB1C3</li>
+
<li>Linearized pSB1C3
 
</ul>
 
</ul>
−
<li>Miniprep B0034</li>
+
<li>Miniprep B0034
−
<li>Transformation in DH5α of interlab studies plasmids</li>
+
<li>Transformation in DH5α of interlab studies plasmids
 +
 
 
</ul>
 
</ul>
 +
 +
<h5>Thursday, June 23th</h5>
 +
<ul>
 +
<li>Digestion E/P pSB1C3
 +
<li>Purification pSB1C3 purified linearized 23/06
 +
<li>Digestion E/P of DesA, DesB, DesC, DesD
 +
<li>3A Assembly of B0034 and I0500 in pSB1K3
 +
<li>Digestion with pSB1C3 23/06 insert digested fliC
 +
<li>Transformation in Tg1 and DH5α
 +
</ul>
 +
 +
<h5>Friday, June 24th</h5>
 +
<ul><li>Digestion E/P pSB1C3, pSB1A3, pSB1K3, pSB1T3, and agarose gel extract
 +
<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>Ligation pSB1C3 and fliC E.Coli / desulfo digested E/P 21/06
 +
<li>Transformation in Tg1 and DH5α
 +
<li>Redone transformation test device 1 and 3 interlab studies
 +
<li>Miniprep pSB1C3 pSB1CA3 pSB1CK3 pSB1CT3
 +
</ul>
 +
 +
<h5>Monday, June 27th</h5>
 +
<ul><li>Transformations DesA, DesB, DesC, DesD, FliC E.Coli and desulfo, interlab studies
 +
<li>Transformations failed
 +
<li>Transformation pSB1T3 miniprep, pSB1K3 miniprep, control
 +
</ul>
 +
 +
<h5>Tuesday, June 28th</h5>
 +
<ul><li>Miniprep promoter+RBS
 +
<li>PCR of IDT sequences (fliC and DesA to D) and gel check
 +
<li>Gel extract pSB1CK3, pSB1CC3, pSB1CT3
 +
</ul>
 +
 +
<h5>Wednesday, June 29th</h5>
 +
<ul><li>Miniprep from interlab studies, T3 and K3
 +
<li>Ligation and transformation in Tg1 Sandra Des A to D, fliC coli desulfo
 +
<li>Redone PCR of IDT sequences (fliC and DesA to D) (different Tm) and gel extract
 +
<li>Transformation of fliCs from remaining ligation in Tg1 Sandra
 +
</ul>
 +
 +
<h5>Thursday, June 30th</h5>
 +
<ul><li>Migration FliC coli, desulfo, DesB, DesC
 +
</ul>
 +
 +
<h5>Friday, July 1st</h5>
 +
<ul><li>Digestion PCR products and vector pSB1C3
 +
<li>PCR clean up on digested pSB1C3, fliC coli desulfo, DesB and DesC, migration and transformation in Tg1
 +
</ul>
 +
 +
<h5>Monday, July 4th</h5>
 +
<ul><li>Isolated white colonies on dishes
 +
</ul>
 +
 +
<h5>Tuesday, July 5th</h5>
 +
<ul><li>Reisolated colonies from 04/07
 +
</ul>
 +
 +
 +
<h5>Wednesday, July 6th</h5>
 +
<ul><li>Colony PCR with FLiC E.Coli Desulfo DesB DesC
 +
</ul>
 +
 +
<h5>Thursday, July 7th</h5>
 +
<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>
 +
 
</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