subtractive hybridisation

[Klaus Salger]

Debbie (D.Carter1 at sheffield.ac.uk) wrote:
: I'm trying to set up a subtractive hybridisation system to look at  
: differential gene expression between two tissue types. I would be most 
: grateful for any hints and tips. I'm particularly interested in the idea 
: of using PCR to increase the amount of driver used as there is limited 
: starting tissue. Any ideas will be warmly recieved.
: Thanks.

Have a look at "Identifying differences in mRNA expression by 
representational difference analysis of cDNA", Hubank & Schatz,
NAR Vol.22 No.25 p.5640 (1994).
This method involves multiple hybridizations and amplifications to
end up with fragments of cDNAs present in the tester but not in the driver 
or beeing present in different amounts.
(I didn't yet test it myself.)

Cheers
  Klaus

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Klaus Salger                phone : +49 (0)89 5902 -502
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