DNA extraction from polyacrylamide gels

Errol E.S.Kwan at massey.ac.nz
Thu Aug 24 20:24:50 EST 2000


I silver stain my Polyacrylamide gels and the acetic acid stop
solution doesn't pose any problem for me.  When I excise my band I
place it in 50 microlitres and leave it at 4 degrees C overnight.  In
the morning it amplifies nicely.

Errol

On 24 Aug 2000 09:04:23 +0100, Vince.Mulholland at sasa.gov.uk (Vince
Mulholland) wrote:

>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
>
>
>___________________________________________________
>How do you extract DNA from a polyacrylamide gel?=20
>
>
>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 =D73 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.=20
>
>
>Thank you=20
>Lise Larsen=20
>
>
>
>
>
>---
>
>
>---






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