priming pcr using wrong species sequences

Laura L. M. Hoopes hoopes at oxy.edu
Sun Jul 7 22:46:54 EST 1991


I am trying to use Gnebank sequences for human DNA repair genes to prime
RT-PCR reactions in mice, and the only protection from evolution I've
incorporated into the experiment so far is to choose regions that are
homolous betwe the proteins I want to study and other known proteins,
hoping they will be the most conserved parts.  The primers are not very
effective and I now want to use primers that are multiple in the third
positions of the codons.  Does anyone have good/bad experience with this
type of experiment?  Any suggestions for length, maximum or minimum amount
of multiple sequences, temperatures?{  Thanx, Laura Hoopes, Hoopes at oxy.edu



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