User:Karmella Haynes/Notebook/BioBrick cloning/2010/03/31

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03/31/10

 * &#x2713; Minipreps: KAH86 re-built (4)
 * &#x2713; Assembly: KAH96 re-build
 * &#x2713; Sequencing (same-day): KAH86 plasmids
 * &#x2713; Cultures: KAH014 (DPRE) (4)

Minipreps > Check with E/P digests > Compare to KAH81

--> Samples 1 and 2 appear to run larger than KAH81. Check all 4 via sequencing and proceed with KAH96 assembly.

Assemblies
 * 1) KAH96: KMC01(5xGal4)/(S/P)/~3300 &#x2713; + (1) KAH86/(X/P)/2726

> Digests (Fermentas FD) --> X/P/ScaI triple digest

> Measure conc.'s

--> KMC01(S/P) = 62.3 ng/μL

> Ligations

--> R.T./ ~10 min.; Add 50 μL Z-DH5α

Sequencing (Genewiz, same-day, 04/01/10) --> Confirm HSVtkTATA-K
 * 1) KAH86-1 P0001: good
 * 2) KAH86-2 P0001: good
 * 3) KAH86-3 P0001: good
 * 4) KAH86-4 P0001: good


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