User:Meng Xiao He/Notebook/fall08/2008/10/31

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Update BsmBI digest did not over digest (band sizes still correct) T4 ligation for 15 minutes made DNA smear into large (8-10Kb) region, so over ligated Next repeat PCR fragments over using existing fragments purify>dephosphorylate fragments>purify>digest with BsmBI>ligate for 1-5 minutes>purify>XmaI digest try crossover PCR PCRs: 1-6: cbb, cco, Duo; 3 then 5 Xover: cbb, cco, Duo