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Team:Paris/6 August 2009
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| - | ==Lab work== | + | ===Lab work=== |
| - | + | ====Microscope==== | |
| - | <div | + | <div class="caroline"> |
| - | FM4-64 dying MG4 and Kayo | + | FM4-64 dying MG4 and Kayo delete Tol R |
| - | </div><div | + | </div> |
| + | <div class="experience"> | ||
protocol n°2 (05/08/09) but using PBS | protocol n°2 (05/08/09) but using PBS | ||
| Line 28: | Line 29: | ||
->not enough dye | ->not enough dye | ||
</div> | </div> | ||
| - | + | ====Molecular Biology==== | |
| - | <div | + | <div class="vicard"> |
Gel Migration | Gel Migration | ||
| - | </div><div | + | </div> |
| - | *[Ladder 1kb| | + | <div class="experience"> |
| - | ** | + | *[Ladder 1kb|[https://2009.igem.org/Team:Paris/Freezer_PCR_products#A1 A1]|nothing|[https://2009.igem.org/Team:Paris/Freezer_PCR_products#A2 A2]|nothing|[https://2009.igem.org/Team:Paris/Freezer_PCR_products#A3 A3]]using 1% agarose during 30min |
| - | ** | + | **[https://2009.igem.org/Team:Paris/Freezer_PCR_products#A1 A1] = Tg3p = 274bp (utiliser le ladder 100bp) |
| - | ** | + | **[https://2009.igem.org/Team:Paris/Freezer_PCR_products#A2 A2] = TE3 = 1036bp |
| - | + | **[https://2009.igem.org/Team:Paris/Freezer_PCR_products#A3 A3] = ClyA (with RBS and poly G linker = 987bp | |
| + | |||
| + | |||
<center>[[Image:KR000634.JPG]][[Image:A3_cut.JPG]]</center> | <center>[[Image:KR000634.JPG]][[Image:A3_cut.JPG]]</center> | ||
| - | * | + | |
| - | **[Ladder 1kb|nothing| | + | |
| - | ** | + | *[https://2009.igem.org/Team:Paris/Freezer_PCR_products#A1 A1] and [https://2009.igem.org/Team:Paris/Freezer_PCR_products#A2 A2] didn't work we don't see it on the gel because it's not in E.Coli. |
| + | *[https://2009.igem.org/Team:Paris/Freezer_PCR_products#A3 A3] didn't migrate very well | ||
| + | **[Ladder 1kb|nothing|[https://2009.igem.org/Team:Paris/Freezer_PCR_products#A5 A5]|nothing|[https://2009.igem.org/Team:Paris/Freezer_PCR_products#A5 A5]|nothing] using 1% agarose during 30min | ||
| + | **[https://2009.igem.org/Team:Paris/Freezer_PCR_products#A5 A5] = TolRII = 279bp | ||
| + | |||
| + | |||
<center>[[Image:KR000637.JPG]][[Image:A5_coupe.JPG]]</center> | <center>[[Image:KR000637.JPG]][[Image:A5_coupe.JPG]]</center> | ||
</div> | </div> | ||
| - | <div | + | <div class="vicard"> |
Purification and Digestion | Purification and Digestion | ||
| - | </div><div | + | </div> |
| - | *Purification of | + | <div class="experience"> |
| + | *Purification of [https://2009.igem.org/Team:Paris/Freezer_PCR_products#A3 A3], [https://2009.igem.org/Team:Paris/Freezer_PCR_products#A4 A4] directly in micro-column, [https://2009.igem.org/Team:Paris/Freezer_PCR_products#A5 A5], [https://2009.igem.org/Team:Paris/Freezer_PCR_products#A6 A6] using gel then micro-column | ||
*Digestion with XbaI and PSTI | *Digestion with XbaI and PSTI | ||
</div> | </div> | ||
| - | + | <div class="guillaume"> | |
| - | * | + | New Miniprep |
| - | *ON | + | </div> |
| - | *Digestion P1 | + | <div class="experience"> |
| - | * | + | *New Miniprep for [https://2009.igem.org/Team:Paris/Freezer_Plasmids#P1 P1]: pSB2K3, [https://2009.igem.org/Team:Paris/Freezer_Plasmids#P13 P13]: pSB1A3, [https://2009.igem.org/Team:Paris/Freezer_Plasmids#P14 P14]: BBa_K136050 (RBS tetR) |
| - | *Transformation 1L7 | + | *Result: Just good for trash. Make new ON culture. |
| + | </div> | ||
| + | <div class="guillaume"> | ||
| + | Digestion by XbaI/PstI (XP) or XbaI/SpeI (XS) | ||
| + | </div> | ||
| + | <div class="experience"> | ||
| + | *[https://2009.igem.org/Team:Paris/Freezer_Plasmids#P1 P1]: pSB2K3 by XP | ||
| + | *[https://2009.igem.org/Team:Paris/Freezer_Plasmids#P2 P2]: BBa_J61002 (plasmid backbone with RFP) by XS | ||
| + | *[https://2009.igem.org/Team:Paris/Freezer_Plasmids#P7 P7]: Bba_R0040 (ptet) by XP | ||
| + | Digestion during 2h at 37°C. | ||
| + | </div> | ||
| + | <div class="guillaume"> | ||
| + | Gel migration | ||
| + | </div> | ||
| + | <div class="experience"> | ||
| + | *Gel migration for [https://2009.igem.org/Team:Paris/Freezer_Plasmids#P1 P1]: pSB2K3, [https://2009.igem.org/Team:Paris/Freezer_Plasmids#P2 P2]: BBa_J61002 (plasmid backbone with RFP), [https://2009.igem.org/Team:Paris/Freezer_Plasmids#P7 P7]: BBa_R0040 (ptet) | ||
| + | *[1kb|P1|P2|P7|100bp|/] | ||
| + | |||
| + | <center>[[Image:Wikidigestion060809.png]]</center> | ||
| + | |||
| + | *Result: | ||
| + | **pSB2K3: wrong bande. Try again... | ||
| + | **plasmid backbone with RFP: Wrong enzymes?! | ||
| + | **ptet: good but the 1500bp band must not be there! | ||
| + | </div> | ||
| + | <div class="guillaume"> | ||
| + | ON culture | ||
| + | </div> | ||
| + | <div class="experience"> | ||
| + | *ON culture for [https://2009.igem.org/Team:Paris/Freezer_Strains_(Glycerol_stock)#S8 S8]: pSB2K3, [https://2009.igem.org/Team:Paris/Freezer_Strains_(Glycerol_stock)#S29 S29]: pSB1A3, [https://2009.igem.org/Team:Paris/Freezer_Strains_(Glycerol_stock)#S30 S30]: BBa_K136050 (RBS tetR) in order to done new miniprep | ||
| + | </div> | ||
| + | <div class="guillaume"> | ||
| + | Transformation | ||
| + | </div> | ||
| + | <div class="experience"> | ||
| + | *Resuspention of biobrick BBa_J2479 (1L7) | ||
| + | *Transformation in competent DH5α | ||
| + | </div> | ||
<html> | <html> | ||
</div> | </div> | ||
| Line 64: | Line 110: | ||
</html> | </html> | ||
| - | ==To do list== | + | ===To do list=== |
{| | {| | ||
|- style="background: #ccccff; text-align: center;" | |- style="background: #ccccff; text-align: center;" | ||
| Line 102: | Line 148: | ||
|Sylvain | |Sylvain | ||
|if(!oligo){return Maltose;} return PCR; | |if(!oligo){return Maltose;} return PCR; | ||
| - | |- style="background: | + | |- style="background:green; text-align: center;" |
|Guillaume | |Guillaume | ||
| - | | | + | |Lab |
|} | |} | ||
{{Paris2009_Calendar_Link|5_August_2009|7_August_2009}} | {{Paris2009_Calendar_Link|5_August_2009|7_August_2009}} | ||
Latest revision as of 09:22, 13 August 2009
Contents |
NoteBook
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Lab work
Microscope
FM4-64 dying MG4 and Kayo delete Tol R
protocol n°2 (05/08/09) but using PBS
->nearly the same as MgSO4
protocol n°2 (05/08/09)but 1ml of cells and 1µl of dye, one with MgSO4 and one with PBS
->not enough dye
Molecular Biology
Gel Migration
- A1 and A2 didn't work we don't see it on the gel because it's not in E.Coli.
- A3 didn't migrate very well
Purification and Digestion
New Miniprep
Digestion by XbaI/PstI (XP) or XbaI/SpeI (XS)
Digestion during 2h at 37°C.
Gel migration
- Gel migration for P1: pSB2K3, P2: BBa_J61002 (plasmid backbone with RFP), P7: BBa_R0040 (ptet)
- [1kb|P1|P2|P7|100bp|/]
- Result:
- pSB2K3: wrong bande. Try again...
- plasmid backbone with RFP: Wrong enzymes?!
- ptet: good but the 1500bp band must not be there!
ON culture
Transformation
- Resuspention of biobrick BBa_J2479 (1L7)
- Transformation in competent DH5α
To do list
| Matricule | TODO |
| Luc | Labs |
| Romain | Look for his feet |
| Charlotte | Jun/Fos system (oligo + system) |
| Stoff | DataBase / kitchen system ? |
| Chris | modeling: model genetique network / next week lab planning / gillepsy |
| Lisa | WTF !!! |
| Caroline | lab / microscope |
| Souf | lab / oligo / wiki |
| Vicard | Lab : Gel :D |
| Pierre | Tol/pal modeling |
| Sylvain | if(!oligo){return Maltose;} return PCR; |
| Guillaume | Lab |
