Northern with non-radioactively labelled probe

Max m.c.lintern at bham.ac.uk
Mon Apr 10 04:20:50 EST 1995


In article <v01510100aba049c73356@[143.48.11.21]>, lodhi at CSHL.ORG
(Muhammad Lodhi) wrote:

> I am wondering if anyone does northern with non-radioactively labeled
> probes (chemiluminiscent, fluorecein or others).  Which system is generally
> used and how different is the sensitivity of signal compared to labelling
> with P32.  Any comments or references will be appreciated.  Thanks
> 
> Muhammad A. Lodhi

Dear Muhammad

I am trying to use cold NB to look at levels of AChE mRNA. I have used a
kit to make a probe with a fluorescein molecule attached. The idea then is
to use an antibody to the fluor. which is alk phos conjugated and develop
to give a coloured band. I have had this kit for over a year and while the
control experimnets it suggests work fine I have a great deal of trouble
getting it to hybridise to an unlabelled peice of probe and have never got
it to pick out any mRNA!!!! This might be due to probs with my probe coz I
know of others using similar kits which have had more succesful results.
Certainly the method seems less sensitive, but if you get anywhere please
let me know!!!!

Maxine Lintern

M.C.Lintern at bham.ac.uk



More information about the Methods mailing list