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
>>>>>>---
>>>---
