Quick/Express/Easy/Etc Hybridization buffers?

Eric Lader elader at ambion.com
Thu Oct 7 21:02:14 EST 1999


In article <071019991212099991%jpcd100 at mole.bio.cam.ac.uk>,
  jpcd100 at mole.bio.cam.ac.uk wrote:
> Hi All, can anyone give me any tips as to the true efficacy of all
> these fancy hybridization buffers being advertised recently. Ambion
had
> an advert in Nature last week showing a northern hybed in "standard"
> buffer and the same blot using their buffer, showing a massive
increase
> in signal.
>
> Are any really as good as they say they are? Thanks for any info or
> recommendations.
>


Hi John,

Lots of people have asked these questions lately. Yes, the data is real.
(but of course, only the best pic makes it into the ad) We see 20X-100X
the signal. Now a couple of fine points.... First, this level of signal
enhancement is true for double stranded DNA probes, not riboprobes -
there we see a smaller increase, usually 2-10X. Second, the increase is
true if you use 10E6 of a high specific activity probe (which is what we
recommend for Ultrahyb). You can jam much more counts in an
unaccelerated/unenhanced hyb mix - 10E7 is possible and you'll get 10X
the signal level (or lots of background...tough to predict).

What we see with Ultrahyb and DNA probes is hyb levels equivalent to
close to 100% target hybridized (like in an ribonuclease protection
assay).

Hey - the samples are free, call and give it  a try.

Eric Lader
--
Senior Scientist
Ambion, Inc.
Austin, Texas


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