ARMS/allele-specific PCR

taylorgr at delphi.com taylorgr at delphi.com
Sat Jan 7 09:20:35 EST 1995


Mike Morris <mike at medsun.unige.ch> writes:
 
>  What are the criteria for designing allele-specific primers? Can 
>anybody point me at a review? Or give their experience?
>  thanks in advance,
>	Mike
 
Mike
Designing ARMS primers is not a pushover. I heard romours the Cellmark had
several attempts to get their CFTR set working. So wherever possible copy
someone elses. It seems to be usual practice to include an additional mismatch
at base 3'-3 as well as the competing bases at the 3' position. Ferrie et al
wrote up the details of the CFTR stuff in Am J Hum Genet, sorry can't remember
the
full ref. They used lower (100uM) than usual conncs of dNTP, less Taq and
low concs of target DNA to enhance specificity. They also reported that
multplex ARMS tests seemed more robust (i.e. less prone to false amping) than
single reaction tests. In my lab we are currently setting up fluorescent ARMS
tests using competing primers of different colours. It requires quite a lot
of adjustment before you hit specific conditions. Good luck.
Graham Taylor, Clinical Molecular Genetics Lab, Leeds UK e-mail
taylorgr at delphi.com



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