To avoid doing many steps of site-directed mutagenesis, we want to PCR
and clone a fragement using one 80mer primer. We have previously used a 50mer,
with no problems, but we cannot amplify with the 80mer. We have tried both from
genomic DNA and from a larger PCR fragment (made with conventional primers).
Does anybody have any tricks for amplifying? We have tried a range of Mg
concentrations, but have never seen any suggestion of a band.
Please answer by email - I don't regularly follow this group.
Medical Genetics, Geneva, Switzerlan