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Denaturing Gradient Gel Electrophoresis

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#1 LauraB



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Posted 07 July 2009 - 01:11 PM


I've been having a problem with my DGGE gels since I started running them. All the lanes have streaks of bands, even when I've run pure cultures. No matter what samples I load they all look the same, leading me to believe it's the gel, not the samples. I use new buffer every 2-3 runs, I have new reagents, the equipment is fairly new. I run the sample for 15 min at 30V to concentrate the DNA in the well and then at 130V for 3-3.5 hours. It's an 8% gel and a 30-55% gradient. Any suggestions would be greatly appreciated. I attached a file of a five wells containing a pure culture and the left was a combination of all five, my attempt at a DGGE marker.

Thanks, Laura

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