User:Tk/Notebook/MF/2008/11/05

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 * style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]] Mesoplasma florum simplification
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 * style="background-color: #F2F2F2" align="center"|  |Main Page


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Vector backbone

 * gave up on biotin purification
 * PCR cleanup after PCR with plain primers (62 C anneal) 100 ul rxn
 * elute 2x with 40 ul in EB
 * digestion in 100 ul with E/P 2 hr / 20 min heat kill
 * PCR cleanup, elute 2x with 40 ul
 * yields 1AC3: 48 1AK3: 69 1AT3: 57 ng/ul

Transformation results

 * one colony on the AC plate, none on others.
 * control normal at 4e7 efficiency
 * culture the colony in Amp/Kan and Amp/Tet medium


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