DGGE technique - staining and general (Apr/22/2002 )

Hi everyone,
people in my division are using DGGE to separate out PCR sequences after using degenerate primers. The silver staining technique we use is not optimal, nor has ethidium bromide been all that good. Any suggestions on using silver staining and DGGE in general ?

-BJSAu-

hi

Are you getting any visible bands using silver staining? I got bands by staining with ethidium bromide for 10 mins, but am having no luck using silver stain

-taggart-

SYBR green works much better than Ethidium bromide for me, I have a mixed template from soil, with degenerate primers as well. It is much more sensitive - I let it stain for 40 minutes.

-Nelson Lab-

Hi

I have started to use sybr gold to stain my gels, I stain them for about 15mins this gives a good result, it is also possible to reuse the stain

-taggart-

I got bands by staining with silver, very well

-Dnase-

I just tried Sybr Gold.
It's working pretty well.
Patrick

-Patrick Laplante-

QUOTE (Patrick Laplante @ Oct 29 2004, 09:58 AM)

I just tried Sybr Gold.
It's working pretty well.
Patrick

Hasvanyone used bio rad silver stain for their gels? If so how successful was it and have you any tips?

-taggart-

I used Ethidium Bromide. I stain for 20 min and it work very good for me and people in our lab.

-freshman-uk-

We use Sybr gold at the moment but want to try silver staining with the bio rad kit but so far we have had no success with this

-taggart-

I used both silver and etbr staining. silver staining for me work much better then etbr. Soon i'll try stain the gel with syber green.

-awanis-