Sensitive stain for DNA in gels

M. Hertel hippocampus at NoSpam.gmx.ch
Fri Mar 23 02:50:09 EST 2001


Mike Brisco wrote:

> What is the most sensitive stain out there for detecting DNA in gels?
> We've tried SYBR green I and II and Gelstar, but need to go more sensitive
> still, preferably without using radioactivity.
>
> Regards,
>
> Mike Brisco.
> Michael J Brisco, PhD
> Department of Haematology & Genetic Pathology
> Flinders Medical Centre
> Bedford Park
> South Australia 5042
>
> tel +61 8 8204 4105
> fax +61 8 8204 5450

> Silverstain of DNA in Acrylamid/Bisacrylamid gels:

I used Minigels 7.5 to 12 %

- prestain fixation 3 min 10 EtOH, 0,5% acetic acid
- 5 min fixation 10% EtOH, 0,5% acetic acid, 0,2 % AgNO3
- 2 min Water
- development at least 5 min (could need much longer)3,0% NaOH (W/V) 0,2%
AgNO3
- 5 min 10%EtOH 0,5% acetic acid
- 10 min Water

You have to play around to reduce the background, I used this method Years
ago to detect a very low band...
The method is described by Cainis and Murray in BioTechniques Vol 17 No. 5
(1994)
Moritz








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