Opinion poll: Clonetech Matchmaker libraries
Simon Dawson
Simon.Dawson at nott.ac.uk
Fri Mar 8 10:19:36 EST 1996
Hi,
Yup. I have used a MatchMaker cDNA library. It was an adult human brain
library and it's worked (touch wood!) fine. As for quality, it depends what you
want. For this assay it may be advantagous to have a 'bad' library in any case
i.e. one in which a proportion of clones are partial. The reason? As I see it
(and I'm probably wrong!), expressing partial sequences (i.e. individual
domains of proteins maybe?) would throw up interactions which you otherwise
miss because the interacting domains are buried within the intact protein. You
may also get more false positives I guess....but what the hell. You'll have to
investigate the biological relevance of your two-hybrid result anyhow, whatever
the libraries quality.
The library I used (cant find the cat. No. just now) was primed with a
mixture of oligo(dT) and pdN6 (random primers), so I am not altogether
surprised that some of the clones I have are partial.
I have however heard from people in our department that ClonTech libraries in
general have a bad reputation. I'm just going to go and pull a full length
clone out of another library :)
Hope thats of some help,
Simon.
--
Dr. Simon Dawson TEL:+44 (0)115 9249924 ex. 44787
Dept. of Biochemistry FAX:+44 (0)115 9422225
Queens Medical Centre Email:Simon.Dawson at .nott.ac.uk
Clifton Boulevard http://www.ccc.nottingham.ac.uk/~mbzspd/Simon.html
Nottingham
U.K. "Back off man, I'm a scientist!" - Bill Murray.
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