allele-specific PCR

Mr. T. Weissensteiner tweissen at hgmp.mrc.ac.uk
Thu Mar 11 12:04:43 EST 1999


Gregor Bucher wrote:
>
> Hi!
> Does anybody has experience with allele-specific PCR. I´d like to have
> some hints how the primers and conditions should be chosen before I start
> some hints how the primers and conditions should be chosen before I start
> to find out  these things again. To my knowledge only A-G C-G and A-A
> mismaches will give results.
>
> What else can I do to make the reaction work?
>
> Thanks a lot!
>
> Gregor

Assuming what you want to develop is a typing system, coamplifying an internal 
control is a must - not only to be sure whether your individual sample has worked,
but it also does wonders for the target specificity! 
(Although any :-) single mismatched allele will amplify given enough cycles, the 
control amplimer will amplify faster and exhaust the available reagents first).



Thomas Weissensteiner
Autoimmunity Group
Jenner Institute for Vaccine Research
Compton / Newbury
Berkshire RG20 7NN

T    :  0044 1635 577920
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email : tweissen at RETURNMEhgmp.mrc.ac.uk
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