DNA pooling.

Beth Wapelhorst weazel at animal.blarg.net
Mon Apr 27 22:33:49 EST 1998


blech!!! I wouldn't use quantitative PCR to do something that's easily
done conventionally!

Also, my experience with microsatellites has shown me that different sized
alleles amplify...differently. Assuming you're using microsatellites,
you'll have to take this into account, because you're likely to find that
smaller alleles amplify more readily than large ones - and if you're
pooling samples, you might lose the larger alleles all together because of
the "competition" - the smaller alleles (even if they're only smaller by a
few nucleotides) will have the amplification advantage.

sugar beth
DNA Hacker's Resource
www.blarg.net/~weazel/DHR/dhr.htm

Yankit Tam <yktam at aecom.yu.edu> writes:

>Hi everyone,

>I am working on a DNA pooling experiment in which I
>pooled DNA from a number of individuals and then a 
>polymorphic marker is amplified via PCR.  The genotypes
>of these individuals are known.  My objective is to
>see if I can get the same allele frequency distribution
>from a single PCR with pooled DNA as I do PCR with 
>individual samples.  Besides fluorescent primers,
>has anyone tried other methodologies to quantiate
>the pcr bands?  I'd be grateful if you could share
>your experience.  Thank you. 

>Kit

>Kit Tam
>Department of Microbiology and Immunology
>Albert Einstein College of Medicine
>Bronx  NY 10461
>USA

-- 
sugar beth wapelhorst              "I don't think I'm alone when I say I'd
weazel at blarg.net                    like to see more and more planets fall
www.blarg.net/~weazel               under the ruthless domination of our
                                    solar system." - Jack Handey



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