Non-radioactive labelling of DNA

Susanne Rohrer "srohrer(removethis)" at immv.unizh.ch
Thu May 4 10:41:15 EST 2000



"D. Durnford" wrote:

> I would like to limit the use of 32P in my lab and I am shopping for
> non-radioactive labelling methods (random prime). I am primarily
> interested a system that is successful with Northern blot
> Hybridizations. I would like to know if anyone would recommend a
> particular system/method. I am concerned with stripping and
> rehybridization, signal detection etc.
>
> Thanks in advance.
>

DIG labeling (Roche) works nicely: you can random label or use PCR
probes, there is a kit for both. There is even one to chemically label
probes-I haven't tried it yet. Southerns can be stripped, and with
Notherns we haven't had great results but that's got to do with RNA, not
the Dig system. Unfortunately it's not cheap. but then again you don't
have the radioactive mess.

there have been lots of posts discussing the method, and the standard
one in the Roche DIG manual works.

--
Susanne Rohrer
Institute of Medical Microbiology
University of Zurich
Switzerland

srohrer at immv dot unizh dot ch






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