User:Karmella Haynes/Notebook/BioBrick cloning/2011/01/20

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01/20/11

 * &#x2713; Cultures & minipreps: KAH179, 180, KAH176-178/MV8 (2 each); 5 mL ea./ 6 hrs.
 * &#x2713; pSB31 vector: for Pc-TF ES cell expression; diagnostic digest & retransformation

Minipreps --> KAH179, 180: E/P/SacI --> KAH176-178/MV8 fwd: E/ApaLI (*mCherry has an ApaLI site) --> KAH176-178/MV8 rev: E/S

--> Note: ***5xGal4 contains ScaI site, thus observed band size should be 2221 bp.

pBS31 vector --> Collab. w/ Manching Ku (Bernstein lab); Use pBS31-RBGpA vector for FRT integration and dox-induced expression of Pc-TF in ES cells --> Vector contains EcoR1 cloning site and NotI site elsewhere --> Do diagnostic digest to check for XbaI, SpeI

--> Conclusion: Cannot use XbaI or SpeI for cloning in this vector. Blunt the vector and the insert.


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