(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:
>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.
Boehringer do a guide that goes with their long PCR kit. Try them.
National Centre for Medical Genetics
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