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文献和实验TOP10 chemically competent cells
reduces transformation efficiency by about 3x the first time, and about 6x total after several freeze/thaw cycles. Measurement of competence Transform 50 μl of cells with 1 μl of standard pUC19 plasmid (Invitrogen) This is at 10 pg/μl
Recombineering/Lambda red-mediated gene replacement
in 2 mL LB + 50 μg/mL Amp add 50 μL 10 mg/mL Ampicillin stock Include enough samples for two conditions: +/- L-arabinose induction When OD600 of cells(+pKD46) reaches 0.1, add L-arabinose to concentration
(10 –19 ).
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