Is Boehringer Mannheim the only supplier of Dig-labellin
nash at nrcbsa.bio.nrc.ca
Fri Jan 20 09:27:09 EST 1995
In article <71724191105991.503674 at usa>, Joseph Polli <polli~j at glaxo.com> wrote:
<prior DIG vs biotin posts deleted>
>Actually, we've obtained excellent results with biotin-labelling
>for most of our applications (Southern blots, using PCR to
>incorporate label, biotinylated oligos) except when performing
>**colony lifts**. We posted this problem a while back to no
>it is kind of a pain. 2) if background is due to bacterial
>colonies, have you tried blocking filters with free biotin, then
>wash, and then block with strepavidin, followed by another wash?
>I hope this helps.
Q: Doesn't E. coli contain heaps of free biotin? That's what I've
been told, anyway - but that's in the realms of hearsay.
DIG is used quite a bit here at NRC for colony lifts, even after
site-specific mutagenesis, with nary a background problem. My
esteemed colleague, Warren Wakarchuk (who is probably lurking here),
swears by it.
John Nash Institute for Biological Sciences *
Email: nash at nrcbsa.bio.nrc.ca National Research Council of Canada * .*
World Wide Web: http://cansnd.cisti.nrc.ca/~nash/home.html *
Disclaimer: Opinions are mine, not NRC's
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