Non-isotopic detection of nucleic acids

Klaus Salger salger at wap18.zi.biologie.uni-muenchen.de
Wed Aug 10 15:08:29 EST 1994


Peter Leif Bergsagel (bergsage at helix) wrote:
: We have a summer student who is not licenced to work with radioactivity,
: and would like to use a non-isotopic method for detection of nucleic acids.
: I reviewed the last 4 months postings in the archives and did not find any
: threads that seemed to address the issue. I would be interested to know
: peoples recommendations for:

: 1. Detection of a single copy gene in 10 ug of human genomic DNA

: 2. Detection of an abundant mRNA (>1000 copies/cell) in 10 ug of total RNA

: 3. Screening tertiary plaques, or colonies, or PCR products


: I shall be glad to summarize the response and post it. Endorsements for
: specific commercial products (kits) are encouraged. I thank all
: respondents in advance.

See Engler-Blum et al. Analytical Biochemistry 210, p.235, 1993.
The authors describe a procedure which they say gives higher sensitivity
then radioactive hybridization. They use DIG-labeling.
I found it to work but didn't compare it with radioactive labeling.
1. and 2. should be no problem with this protocol. As far as I remember
3. using DIG-labling was already discussed in this newsgroup (see archive).
If you don't have access to the paper, I could e-mail you the protocol.
Hope this helps
  Klaus

Klaus Salger
Zoologisches Institut
AG MacWilliams
Luisenstr. 14
80333 Muenchen
Germany

phone : ++49 (0)89 5902 -502
FAX   :                 -450
e-mail: salger at zi.biologie.uni-muenchen.de



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