User:Karmella Haynes/Notebook/BioBrick cloning/2010/01/20

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01/20/10

 * &#x2713; PCR: KAH94, 95/JDS48/49 minipreps from 1/19
 * &#x2713; Minipreps: KAH139, 140, 141 (2 each); KAH118/MV2(12)
 * &#x2713; Assemblies: KAH139, 140, 141/ V0201

Minipreps > Check with E/P digests

PCR --> Check KAH94, 95/JDS48/49 minipreps from 1/19 (24 total) --> Primers: RFP1 (used for RFP expression RT-PCR; should also work on DNA)

--> PCR (96-well): 95°C/ 3 min.; [95°C/ 30 sec., 57°C/ 30 sec., 72°C/ 30 sec] x30; 72°C/ 3 min. --> Still unclear. Tomorrow digest w/ BglII

Assemblies
 * 1) KAH139/V0201: (1) KAH139/(N/S)/2945 + (4) V0201/(N/S)/5157
 * 2) KAH140/V0201: (2) KAH140/(N/S)/2939 + "
 * 3) KAH141/V0201: (3) KAH141/(N/S)/2527 + "

> Digests (Fermentas FD) --> Use ScaI to cut V0120 for KAH139, 140

> Measure conc.'s

> Ligations


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