Help! PCR products cloning

Gabby Gerlitz gabiger at zoot.tau.ac.il
Mon Sep 9 08:07:47 EST 1996


Feng Lin wrote:
> 
> I am cloning DNA fragments amplified by PCR with Taq DNA polymerase.There are BamHI
> sites at each 5'end of my primers and three additional bases(GTG or GCC) out of the
> restriction site as protection bases.The experiments seem that the PCR products cannot
> be digested by incubation with BamHI and cloned in pBS/BamHI.Other enzymes sites at PCR
> primer(eg,PstI)can work well in my lab.The quality of primers may be O.K.But where's the
> problem?
> Thank you for your help!
> 
> Feng Lin

Hi Feng Lin,

It seems that BamH1 needs more then 3 bases at each side of the restriction 
site in order to cut. There are some cataloges which that kind of 
information is writen. Try to check it. If this is the case you will have to 
design new primers with longer tails.

Happy new year(the jewish year comes to an end and to a new start next 
week),
Gabi.



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