SYBR Green I, more exper
junkim at peaplant.biology.yale.edu
Fri Sep 2 19:49:38 EST 1994
In article <199409011917.MAA01885 at netcomsv.netcom.com>,
Jeanne Loring <jloring at genpharm.com> wrote:
>YES!! I inadvertently did the same experiment, with a 3% NuSieve TBE gel
>containing some <100 bp PCR products. I left it over a weekend in 1:10,000 SYBR
>green in water, and on Monday, there was very little diffusion. The DNA in same
>type of gel stained with EtBr would be very diffuse if not gone in that much
>time ( I do this experiment a lot). I assume that SBYR green stabilizes the
>DNA, and remains tightly bound, perhaps changing the conformation or the charge
>of the DNA, making it less diffusible. The literature that comes with the dye
>says that you can prestain the DNA before running it on a gel, and that this
>DOES affect the migration. I assume it affects all DNA similarly, so putting it
>in the gel will just alter the migration of both standards and experimental
>bands equally, and make no difference. However, I don't know whether it binds
>stoichiometrically, or if it EVER comes off, so I've been loathe to try it for
>quantitation or prep gels. Anyone know?
I called up Molecular Probes technical support to specifically ask about this.
The conversation went something like,
Me: How do get this off?
Them: Uh, use standard techniques...
Them: GeneClean should work.
Me: How about n-butanol?
Them: Uh, yeah, that should work too.
Me: Has anybody tried any of this?
Them: Uh....Let me check...I'm not sure.
At this point I thanked them and hung up. I guess one of these days, I'll
try band cutting...
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