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DGGE - DNA migration

Discussion of all aspects of biological molecules, biochemical processes and laboratory procedures in the field.

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DGGE - DNA migration

Postby TomMol » Sat Apr 30, 2005 1:44 pm

Hello,

Two months ago I started use DGGE method. I want to use it to detection bacteria (mainly Lactobacillus) in stool. I have got problem. When I put PCR product into wells samples didn’t migrate equally. It migrate faster on both periphery gel and slower in center. I added 100 ul 10% APS and 10 ul TEMED per 25 ml poliacrylamide. I used 100 V and temperature 60C – 10 % gel.

What should I change in my procedure to my results will be better?
Thank you

Tom

tgosiews@cm-uj.krakow.pl
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Postby Niqqie » Fri May 20, 2005 6:18 pm

Either it is a problem in the way the electricity is flowing through your gel, or it is a problem of how the gel polymerizes. I personally think 100V is a lot, I run 7% polyacrylamide gels at 60V; I only use a high voltage to get the samples into the gel, after which I turn it down.
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