Team:Paris/12 August 2009
From 2009.igem.org
(New page: {{Template:Paris2009}} {{Template:Paris2009_menu}} ==NoteBook== {| style="color:#1b2c8a;background-color:#0;" cellpadding="0" cellspacing="1" border="0" bordercolor="#fff" width="100%" a...) |
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==NoteBook== | ==NoteBook== | ||
- | + | {{Paris2009_Calendar}} | |
- | { | + | {{Paris2009_Calendar_Link|11_August_2009|13_August_2009}} |
- | + | <center> '''August 12th''' </center> | |
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- | + | ||
- | + | ||
- | + | ||
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<html> | <html> | ||
</div> | </div> | ||
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+ | ===Lab work=== | ||
+ | <div class="vicard"> | ||
+ | Ligation | ||
+ | </div> | ||
+ | <div class="experience"> | ||
+ | [https://2009.igem.org/Team:Paris/Freezer_Digestion_Products#D15 D15] (2xterminator E/S) and [https://2009.igem.org/Team:Paris/Freezer_Digestion_Products#D16 D16] (pLac E/X) with vector psb2k3 | ||
- | + | DO : D15=1,19µg/mL D16=1,75µg/mL | |
+ | {| | ||
+ | |- style="background: #b3d0E4; text-align: center; color:black; " | ||
+ | |width=10%| | ||
+ | |width=10%| | ||
+ | |width=10%| vecteur | ||
+ | |width=10%| insert | ||
+ | |width=10%| H20 | ||
+ | |width=10%| Buffer 10x T4 ligase | ||
+ | |width=10%| T4 ligase | ||
+ | |width=10%| Total | ||
+ | |||
+ | |- style="background: #E0E9EF; text-align: center;" | ||
+ | |width=10%| L5 | ||
+ | |width=10%| 2xterm:pSB2K3(with pLac) | ||
+ | |width=10%| 1µl | ||
+ | |width=10%| 3µl | ||
+ | |width=10%| 13µl | ||
+ | |width=10%| 2µl | ||
+ | |width=10%| 1µl | ||
+ | |width=10%| 20µl | ||
+ | |||
+ | |- style="background: #E0E9EF; text-align: center;" | ||
+ | |width=10%| control - | ||
+ | |width=10%| o | ||
+ | |width=10%| 1µl | ||
+ | |width=10%| o | ||
+ | |width=10%| 15µl | ||
+ | |width=10%| 2µl | ||
+ | |width=10%| 1µl | ||
+ | |width=10%| 20µl | ||
+ | |} | ||
+ | |||
+ | |||
+ | 1h Home temperature | ||
- | |||
</div> | </div> | ||
- | <div | + | |
+ | <div class="vicard"> | ||
+ | Transformation | ||
</div> | </div> | ||
- | <div | + | <div class="experience"> |
- | + | classic protocol with DH5a | |
+ | </div> | ||
- | == | + | <div class="vicard"> |
+ | Purification | ||
+ | </div> | ||
+ | <div class="experience"> | ||
+ | *Purification of A10(ClyA) (PCR at 55° during 30sec) (using 1% agarose during 25 min) | ||
+ | *A10=[https://2009.igem.org/Team:Paris/Freezer_PCR_products#Freezer#A10 A10] 987pb | ||
+ | <center>[[Image:ClyA_pcr2.JPG]]</center> | ||
+ | <center>[[Image:Cly1_pcr3.JPG]]</center> | ||
+ | <center>[[Image:ClyA_coupé.JPG]]</center> | ||
+ | *A10 purificated | ||
+ | <center>[[Image:ClyA_purif.JPG]]</center> | ||
+ | *Purification of A10 work | ||
+ | </div> | ||
- | + | <div class="sylvain"> | |
+ | PCR Gel verification | ||
+ | </div> | ||
+ | <div class="experience"> | ||
+ | 1% aga gel after PCR under different conditions for A18 (OmpA-Linker from Varsaw). | ||
+ | Expected weight : 525bp. | ||
+ | <center>[[Image:Gel_090812Sylvain.JPG]]</center> | ||
+ | Obviously, something has gone wrong. Next step will be different temperature gradients testing for PCR and/or new matrix. | ||
+ | </div> | ||
+ | <div class="sylvain"> | ||
+ | Transformation | ||
+ | </div> | ||
+ | <div class="experience"> | ||
+ | Classic protocol with DH5a, and pSB2K3 from iGEM DNA plates (1/7C). ON culture on LB + Kan. | ||
+ | |||
+ | To be checked tomorrow ! | ||
+ | </div> | ||
+ | |||
+ | <div class="sylvain"> | ||
+ | ON culture for Minipreps tomorrow | ||
+ | </div> | ||
+ | <div class="experience"> | ||
+ | pSB1A3, pLAC, RBSTetR, OmpA-L, Three ligation clones, ON culture in LB + Abiotics. 37°C with stiring. | ||
+ | </div> | ||
+ | |||
+ | <div class="sylvain"> | ||
+ | ON PCR | ||
+ | </div> | ||
+ | <div class="experience"> | ||
+ | A20 : 49, 51 and 53°C PCR with the plasmid for OmpA-L. Overnigth. 15s elongation time. | ||
+ | </div> | ||
+ | |||
+ | <div class="guillaume"> | ||
+ | All Manip | ||
+ | </div> | ||
+ | <div class="experience"> | ||
+ | Comming soon | ||
+ | </div> | ||
<html> | <html> | ||
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</html> | </html> | ||
+ | ===To do list=== | ||
+ | |||
+ | {| | ||
+ | |- style="background: #ccccff; text-align: center;" | ||
+ | |width=100px| Matricule | ||
+ | |width=800px| TODO | ||
+ | |- style="background: #59eb26;text-align: center;" | ||
+ | |Luc | ||
+ | | | ||
+ | |- style="background: #efe715;text-align: center;" | ||
+ | |Romain | ||
+ | | | ||
+ | |- style="background: #d8279c;text-align: center;" | ||
+ | |Charlotte | ||
+ | | | ||
+ | |- style="background: #fbfbfb;text-align: center;" | ||
+ | |Stoff | ||
+ | | | ||
+ | |- style="background: #0824f5;text-align: center;" | ||
+ | |Chris | ||
+ | | | ||
+ | |- style="background: #06ff00;text-align: center;" | ||
+ | |Lisa | ||
+ | | | ||
+ | |- style="background: #b40ecd;text-align: center;" | ||
+ | |Caroline | ||
+ | | | ||
+ | |- style="background: #ff9966;text-align: center;" | ||
+ | |Souf | ||
+ | | | ||
+ | |- style="background: #79f4f8;text-align: center;" | ||
+ | |Vicard | ||
+ | | | ||
+ | |- style="background: #e35050;text-align: center;" | ||
+ | |Pierre | ||
+ | | | ||
+ | |- style="background: #bababa;text-align: center;" | ||
+ | |Sylvain | ||
+ | |pSB2K3 transformation ; PCR gels and purifs or new culture for minipreps | ||
+ | |- style="background: #009900;text-align: center;" | ||
+ | |Guillaume | ||
+ | | | ||
+ | |} | ||
- | |||
- | + | {{Paris2009_Calendar_Link|11_August_2009|13_August_2009}} |
Latest revision as of 22:37, 16 August 2009
NoteBook
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Lab work
Ligation
D15 (2xterminator E/S) and D16 (pLac E/X) with vector psb2k3
DO : D15=1,19µg/mL D16=1,75µg/mL
vecteur | insert | H20 | Buffer 10x T4 ligase | T4 ligase | Total | ||
L5 | 2xterm:pSB2K3(with pLac) | 1µl | 3µl | 13µl | 2µl | 1µl | 20µl |
control - | o | 1µl | o | 15µl | 2µl | 1µl | 20µl |
1h Home temperature
Transformation
classic protocol with DH5a
Purification
- Purification of A10(ClyA) (PCR at 55° during 30sec) (using 1% agarose during 25 min)
- A10=A10 987pb
- A10 purificated
- Purification of A10 work
PCR Gel verification
1% aga gel after PCR under different conditions for A18 (OmpA-Linker from Varsaw). Expected weight : 525bp.
Obviously, something has gone wrong. Next step will be different temperature gradients testing for PCR and/or new matrix.
Transformation
Classic protocol with DH5a, and pSB2K3 from iGEM DNA plates (1/7C). ON culture on LB + Kan.
To be checked tomorrow !
ON culture for Minipreps tomorrow
pSB1A3, pLAC, RBSTetR, OmpA-L, Three ligation clones, ON culture in LB + Abiotics. 37°C with stiring.
ON PCR
A20 : 49, 51 and 53°C PCR with the plasmid for OmpA-L. Overnigth. 15s elongation time.
All Manip
Comming soon
To do list
Matricule | TODO |
Luc | |
Romain | |
Charlotte | |
Stoff | |
Chris | |
Lisa | |
Caroline | |
Souf | |
Vicard | |
Pierre | |
Sylvain | pSB2K3 transformation ; PCR gels and purifs or new culture for minipreps |
Guillaume |