PCR using M13 uni & reverse primers
Harry Witchel
Harry.Witchel at Bristol.Ac.Uk
Sun Oct 20 13:59:26 EST 1996
Felicitations Molecular Mages --
I am interested in using the M13 uni and reverse primers to amplify
inserts in vectors which have both Uni and reverse -- I would use entire
bacteria as template (boiling the bugs first). I have tried this with
one set of uni and reverse primers, and it did not work. A heard that
the success of this amplification depends explicitly on precisely which
Uni and reverse primers one chooses. Does anyone have any succesful
experience with this technique? I was going to use:
M13 forward -- 24 bases
CGC CAG GGT TTT CCC AGT CAC GAC
Tm= 73.6°C
m13 rEVERSE -- 23 bases
AGC GGA TAA CAA TTT CAC ACA GG
Tm = 62.3°C
Many thanks,
Harry
Harry.Witchel at Bristol.Ac.Uk
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