kkorth at kcc.com
Thu Jan 18 09:51:26 EST 1996
pnh at cockleberry.ncifcrf.gov (Paul N Hengen) wrote:
>baharvey at swansea.ac.uk wrote:
>: Re the syber green comments last month concerning its cost, shelf life
>: compared to EtBR etc etc. I put the concentrated stock in about 5 mls
>: of loading buffer which I keep at -20'C in the dark (ie the conditions
>: recommended for the stock). That loading buffer/sybergreen batch has
>: lasted for months now, with no loss in activity (Unlike the
>: sybergreen/running buffer staing solution as recommended by the
>: manufacturers, which lasts about two weeks max).
>: Try it and see
>What is the recipe for your loading buffer and the final conc. of SGI?
I have tried SYBR I Green in loading buffer (standard 6X TBE loading
buffer with 1:10,000 SYBR I Green) as recommended by a newgroup poster
about 6 months ago and found the migration to be significantly affected
by the SGI, such that the bands that were supposed to line up, no longer
did (including my ladder DNA). Same thing happened when adding the SGI
to the agarose itself rather than the loading buffer. Though it did work
as a post run stain, it is just too expensive to use that way and really
gave us no better results than the EtBr (as a post run stain).
I love the sensitivity of SGI and the very low background, but I could
not tollerate the way it affected migration. We no longer use it for
KCC Microbiology Research Laboratories
Neenah, WI 54956
E-mail: kkorth at kcc.com
The ideas expressed here are my own and not
necessarily that of my empolyer.
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