Lise,
Rather than your extraction method, it could be to do with the staining
method you use. Do you silver stain your DGGE gels? If you silver stain,
do you use a stop solution which contains acetic acid? The acetic acid,
after silver staining, seems to render DNA refractory to amplification
(presumably due to nick of the strands).
Vince
Vincent Mulholland,
Senior Plant Pathologist,
Diagnostics & Molecular Biology Section,
Scottish Agricultural Science Agency,
East Craigs,
Edinburgh EH12 8NJ, U.K.
URL: http://www.sasa.gov.uk/
E-Mail: Vince.Mulholland at sasa.gov.uk
Tel:+44(131)2448845 Fax:+44(131)2448912
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How do you extract DNA from a polyacrylamide gel?
I run DGGE gels and need to excise the bands for PCR reamplification and
sequencing. I've tried cut out bands and freeze/thaw them ×3 in water
but didn't get any product after the following PCR. Please, let me know
if you know of a good method to recover the DNA.
Thank you
Lise Larsen
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