Dear all
Concernig CG clamps to add at the end of primers for PCR products cloning, the only information I have is that them assist to subsequent cloning. Could anybody explain me in which way?
Thanks a lot
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hennry
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Posted 22 July 2003 - 02:54 AM
Hi,
a little bit late but maybe you're still interested in this question. ;-)
Well, I usually add some GC-pairs at the end of my primers to enhance
cleavage of the PCR-product at the ends with restriction endonucleases
at sites I introduced with the primers. I don't do this if I want to clone the
PCR-product into a TA-vector (because I don't have to cleave it at the ends).
In the catalogue of New England Biolabs there is a short but nice and
informative chapter about the cleavage efficiency of various restriction
endonucleases at the ends of linear DNA-molecules and a lot of more
information about cloning strategies as well. It's worth having a look on it.
I hope I could help a little bit. Good luck.
With best regards,
Henry
a little bit late but maybe you're still interested in this question. ;-)
Well, I usually add some GC-pairs at the end of my primers to enhance
cleavage of the PCR-product at the ends with restriction endonucleases
at sites I introduced with the primers. I don't do this if I want to clone the
PCR-product into a TA-vector (because I don't have to cleave it at the ends).
In the catalogue of New England Biolabs there is a short but nice and
informative chapter about the cleavage efficiency of various restriction
endonucleases at the ends of linear DNA-molecules and a lot of more
information about cloning strategies as well. It's worth having a look on it.
I hope I could help a little bit. Good luck.
With best regards,
Henry
