IGEM:Paris Bettencourt 2012/Notebooks/Semantic group/day by day//2012/07/21: Difference between revisions
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== Migration on 1% agarose gel == | == Migration on 1% agarose gel == | ||
Migration of 10µL of the digestions | |||
[[Image:Gel_19_07_Digestion.png|thumb|center|500px|Agarose gel of the previous digestion. E : EcoRI; P: PstI; X : XbaI; S : SpeI; Ø : no enzyme.]] | |||
The bands are as expected, we can PCR purif the digestion (2 tubes of 30 µL) | |||
== PCR purif == | |||
The PCR purif is made with the promega kit, I recover dna in 30 µL of water. | |||
* pSC001 : 83 ng/µL | |||
* pSC002 : 76 ng/µL | |||
-> good. | |||
== Ligation of pSC002 dig. + pSC003 dig. == | |||
* vector (pSC002) : 1 µL (ie : 76ng) | |||
* insert (pSC003) : 37 µL (*) | |||
* buf. 10X : 1µL | |||
* T4 dna ligase : 1µL | |||
* water : 4 µL | |||
-> 22°C | |||
Revision as of 07:54, 21 July 2012
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Making -20°C & -80°C stocks
Miniprep of : pSC001a, pSC002a&b, pSC003a
->the big amount of lysis solution did not perturb the miniprep. Digestion of pSC001(a) & pSC002(a)
--> 30 min, 37°C water bath. Migration on 1% agarose gelMigration of 10µL of the digestions The bands are as expected, we can PCR purif the digestion (2 tubes of 30 µL) PCR purifThe PCR purif is made with the promega kit, I recover dna in 30 µL of water.
-> good. Ligation of pSC002 dig. + pSC003 dig.
-> 22°C
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