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文献和实验- normally blunt-end ligation of insert to vector does not reform the restriction site (therefore when you design the oligo that make sure it doesn't reform the restriction site when ligated), therefore only religated vector is digested. See one-tube PCR
the restriction enzymes well. g) Keep the buffer in small aliquots as the freezing/defrosting cycles may damage the ATP. Also do not use too high a concentration of ATP (max. 0.5mM) for blunt-end ligation as polyadenation of blunt-ended may occur? h) The ligase
Agar Medium/Liquid Media XL1BMRF' (Stratagene) LB-Tet JM101 M9 ED8767 LB 2. Pick several colonies into a 12 X 75 mm Falcon tube containing a 2 ml aliquot of the respective liquid media, and incubate for 8-10 hours at 37degC
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