Team:Paris/12 August 2009
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<center>[[Image:ClyA_purif.JPG]]</center> | <center>[[Image:ClyA_purif.JPG]]</center> | ||
*Purification of A10 work | *Purification of A10 work | ||
+ | </div> | ||
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+ | <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> | ||
Revision as of 15:58, 12 August 2009
Contents |
NoteBook
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Brain work
edit please ^^
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.
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 |