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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