Purifying small amounts of DNA for PCR

Mikael Niku mikael.niku at helsinki.fi
Tue May 23 03:32:49 EST 2000


Hello!

Which method would you recommend for purifying
small amounts of DNA to be used as a PCR template,
in order to minimize loss of DNA in the procedure?

I'm trying to PCR DNA from microdissected histological
samples, digested with proteinase K. Each sample typically
consists of a piece of tissue (perhaps 100's or 1000's of cells,
and I should be able to detect a single-copy gene present
in only part of them) in 20 ul digestion buffer. My problem is that
using the crude digest as a template, I'm getting annoying unspecific
products. These are not produced when using purified DNA as a template,
so they probably are related to the purity of the template.

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   Mikael Niku             URL: www.helsinki.fi/~mniku/
   University of Helsinki  Dept. Basic Veterinary Sciences
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                                              - Gandhi
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