Difference between revisions of "Team:Paris Bettencourt/Notebook/Phytase"
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<br><h1>13/08/15</h1> | <br><h1>13/08/15</h1> | ||
+ | <h2>PCR purification</h2> | ||
+ | Protocol: | ||
+ | <u>Dilute</u> PCR product (5 or 10 times ?) with the resuspension buffer. | ||
+ | <u>Pour</u> it in a purification column. | ||
+ | <u>Centrifuge</u> 30sec at 14K rpm | ||
+ | <u>Throw</u> the filtrat | ||
+ | <u>Add</u> 700µL of EtOH (washing solution) | ||
+ | <u>Centrifuge</u> 30sec at 14K rpm | ||
+ | <u>DThrow</u> the filtrat | ||
+ | <u>Add</u> 500µLof washing solution | ||
+ | <u>Centrifuge</u> 30sec at 14K rpm | ||
+ | <u>Throw</u> the filtrat | ||
+ | <u>Centrifuge</u> 30sec at 14K rpm | ||
+ | <u>Throw</u> the filtrat | ||
+ | <u>Put</u> the column in a Eppendorf | ||
+ | <u>Add</u> 45µL of RNAse/DNAse free water right on the membrane | ||
+ | <u>Wait</u> 2min | ||
+ | <u>Centrifuge</u> 2min at 10K rpm<br> | ||
+ | <br> | ||
+ | <h2>PCR The PCR control with an electrophoresis</h2><br> | ||
+ | <br> | ||
+ | <img src="https://static.igem.org/mediawiki/2015/d/d8/Paris_Bettencourt_PCR.png" width="700px"> | ||
+ | We expected strips around 1.300bp. | ||
+ | The strip corresponding to marker with FRT is bigger than the two others strips wich have just the Kan resistance with tails.<br> | ||
− | + | <br><h2>pre-culture</h2> | |
+ | Swo one colony saccharomyces (...) of the yesterday in 5mL of the liquid YPD medium, let 's grow overnight.<br> | ||
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+ | <br><h1>14/08/15</h1> | ||
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Revision as of 17:35, 14 August 2015