(Long-Distance) PCR with genomic DNA

Dr David E Barton dbarton at acer.gen.tcd.ie
Tue May 23 15:50:53 EST 1995


In article <3peipl$6ov at nuscc.nus.sg>,
Tay Hou Ngee Agnes <mcbtayhn at leonis.nus.sg> wrote:
>Hi
>I'm trying to do long-distance PCR with genomic DNA as template and 
>having problems. I noticed that the +ve control supplied which works well 
>uses 30mers with high Tm of 70+-80, anyone knows why? I used to do PCR 
>with genomic DNA as template using 20mers with Tm abt 50-60 and worked 
>fine but those were 200bp products. 
>What are the guidelines for primer length, Tm and annealing temp for 
>PCR with genomic DNA in general, and long-distance PCR in particular?
>Advice much appreciated.
>Thanks
>Agnes

Boehringer do a guide that goes with their long PCR kit. Try them.


Dave Barton
National Centre for Medical Genetics
Dublin, Ireland. 



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