User:Karmella Haynes/Notebook/BioBrick cloning/2010/02/19

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02/19/10

 * &#x2713; DPRE cloning: BB PCR
 * &#x2713; Assmebly: DPRE into V0120

PCR --> Template: DPRE from CJW --> Primers: BBp_DPRE/ BBs_DPRE

--> BioRad Block A --> Zymo clean & concentrate; elute w/ 20 μL dH2O
 * 95°C/ 3 min.
 * [95°C/ 1 min., 57°C/ 1 min., 72°C/ 2 min.] x30
 * 72°C/ 3 min.
 * 4°C/ ∞

Assembly
 * Note: DPRE is not a standard BioBrick. Will need to mutate 2 PstI sites to make it a BioBrick. For preliminary in vivo tests, the wild-type form will be used (without PstI cloning)
 * 1) DPRE: (1) DPRE PCR/(E/S)/1821 + (2) V0120/(E/S)/3200

> Digests

> Measure conc.'s

> Ligations


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