Difference between revisions of "Team:Paris Bettencourt/Notebook/Riboswitch"
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<li>Aliquot in 1.5mL eppendorf tubes and store at -80°C until use.</li> | <li>Aliquot in 1.5mL eppendorf tubes and store at -80°C until use.</li> | ||
</ol> | </ol> | ||
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+ | <br><br> | ||
+ | <h1 class="date one">July 30th</h1> | ||
+ | |||
+ | <h2>Goal 1</h2> | ||
+ | PCR my G block and backbone | ||
+ | |||
+ | <h2>Procedure</h2> | ||
+ | |||
+ | <li>PCR1 = Backbone + 068 and 069 primers, before purify</li> | ||
+ | <li>PCR2 = FINAL + 068 and 069 primers, before purify</li> | ||
+ | <li>Then PCR purification</li> | ||
+ | <li>Nanodrop : PCR 1 = 61ng/uL ; PCR2 = 25,4 ng/uL</li> | ||
+ | |||
+ | <h2>Goal 2</h2> | ||
+ | Digestion/Ligation/Transformation | ||
+ | |||
+ | <h2>Procedure</h2> | ||
+ | |||
+ | <li> PCR1 | ||
+ | ADN : 40uL = 2,4 ug | ||
+ | 4uL EcoRI, PstI, FastAp | ||
+ | 12 uL FD buffer | ||
+ | 56 uL water | ||
+ | TOT =120uL</li> | ||
+ | |||
+ | <li> PCR2 | ||
+ | ADN : 40uL = 1ug | ||
+ | 6 uL EcoRI, PstI | ||
+ | 12 uL FD buffer | ||
+ | 64 uL water | ||
+ | TOT= 128uL</li> | ||
+ | |||
+ | <li> 10 min at 37°C</li> | ||
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+ | <li> Then PCR purification : Nanodrop : nanodrop : PCR 1 = 20,7 ng/uL ;PCR2 = 25,7</li> |
Revision as of 16:39, 4 September 2015