User:Karmella Haynes/Notebook/BioBrick cloning/2010/07/27

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07/27/10

 * &#x2713; Assembly: start re-cloning pAct5C/microMCS (from 7/25/10)
 * &#x2713; Cultures/ minipreps: KAH015 (4, 2 mL)

Digests > Mut mCh clones from last week > Check with X/P digests (unique XbaI introduced by microMCS insert) > Will need to confirm proper orientation of microMCS by sequencing

--> Send all 4 to Genewiz for sequencing on Wednesday

Assembly --> re-do pAct5C/microMCS (from 7/25/10)
 * 1) KFV1: (1) pAct5C/(S/N)/~7000 + microMCS/(ΔN/ΔS dsOligo)/~35

> Dephosphorylation (Roche) --> pAct5C/(S/N)

>Kinase rxn. and Oligo annealing --> Need to phosphorylate microMCS before inserting into dp vector (overnight)

7/28/10 > Ligations


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