cDNA library screening
cerbe at mcw.edu
Mon Jun 4 07:57:49 EST 2001
I am screening a rat vestibular (ear tissue) cDNA library and trying to
pull out some full length messages that we're interested in (previously
discovered using RT-PCR). I'm using 40bp overgoes probes and so far all
of my hits have been non-rat homologies (for example 18 days mouse
embryo clone). I'm wondering if this is typical and I just need to
screen more clones, or if I need to use a larger (>100bp) probe. I've
never done library screening before, so I'm not sure what to expect.
Overgoes probes are also new to me. I am getting an average of 2-5 hits
per 20,000 clones.
Is that typical?
The library is normalized and the cDNA's are in pT7T3 phagemid vector.
I'm screening by colony hybridization.
I'd appreciate any input. Thanks,
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