Q:QuickChange and deletions

Hadi hadia at bdg10.niddk.nih.gov.removethis
Wed Jan 7 22:04:58 EST 1998


Hi netters,
I am just wondering if someone has experience in deletions (let's say 100 
bp) using the Stratagene QuickChange kit. I have found that the method is 
working pretty good with point mutations (at least 90% eff.) but for some 
primers it needed some fiddeling around with the temperature profile to 
get it to work. Any suggestions about primer design when doing deletions? 
Stratagene suggests a formula to calculate the Tm where the "percentage 
of mismatch" is taken into account. Do you consider "mismatches" in loop-
out mutations at all? I'd say that there is no mismatch since the primer 
anneals perfectly to the template (but not the other way around...).

Thanks,

Hadi




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