User:Karmella Haynes/Notebook/BioBrick cloning/2009/12/02

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12/02/09

 * &#x2713; Minipreps: KAH011(8); sequencing order
 * &#x2713; Digests: check MV2
 * &#x2713; Assembly: Re-do KAH93, 94, 95 in MV2 (longer MV2 digest, dephos)

Minipreps > Check via sequencing (Genewiz)

Digests > Cloning with MV2 gives high background > Check MV2 w/ EcoRI, PstI and SpeI individually

Note: Looks like all three cut fine. For cloning (E/P), try a longer digest with more enzyme. Also dephos. the cut vector.

Assemblies
 * 1) KAH93/MV2: (1) KAH93/(E/P)/4839 + (4) pcDNA3.1+puro/ΔCMV(MV2)/(E/P)/4529
 * 2) KAH94/MV2: (2) KAH94/(E/P)/5011 + "
 * 3) KAH95/MV2: (3) KAH95/(E/P)/5083 + "

> Digests (Fermentas FD)

> Measure conc.'s

> Dephosphorylation (Roche) --> Do 2 rxn's

> Ligations


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