Cloning GFP into M13 vector?
r.brooks.robey at mcmail.vanderbilt.edu
Thu Oct 19 20:00:17 EST 1995
Your problem may not be specific for GFP but may reflect a more general
problem with subcloning with blunt-ended PCR products: Is your PCR
product generated using Taq polymerase? If so, you might consider
polishing your blunt-ends before cloning into M13, as this has been show
by several investigators to improve ligation efficiency (I can dig up
references if you desire), presumably because Tag has a nasty habit of
tacking on an EXTRA nucleotide residue on the end . . .
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