I have a graduate student who is using RT-PCR to amplify a large section
of the coding region (75%) of a peptide processing enzyme from several
mammalian species. We intend to sequence these and compare the sequences
to identify conserved residues. His committee is concerned about the
fidelity of Taq polymerase even though he is directly sequencing the PCR
products. They have suggested other polymerases or pooling PCR products
obtained under different amplification conditions to "suppress" errors.
What do you PCR jockeys out there suggest to maximize fidelity in our
situation? Thanks for any and all replys.
Associate Professor of Biochemistry