cross-species PCR

Carl Lawyer clawyer at SIUMED.EDU
Sun Dec 18 15:10:13 EST 1994


Dear PCR netters,
We have sequenced a bovine gene and are now trying to get the same in
human. The problem is we found the bovine sequence came out without a
single nt change when we did the human. This was cross-contamination,
because we had worked with bovine cells in the same room.

When redone without cross-contamination, we get nothing.

Any tips on better cross-species primer design?  Are there any good
references on this?

Codon degeneracy, cross-species homolgous amino acid substitutions, and
intra species and intragene codon bias are all problems.

Any practical solutions? (aside from determining the amino acid sequence of
the human protein, a big chore)

Thanks for any help.
        Carl Lawyer M.D., Director of Asthma Center,        clawyer at siumed.edu
        Southern Illinois University School of Medicine
        Division of Pulmonary Medicine
        PO Box 19230
        751 North Rutledge
        Springfield, IL 62794-9230      Fax 217-788-5543





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