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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