2-hybrid libraries

Chris Fisher cfisher1 at chartermi.net
Wed Aug 31 21:35:30 EST 2005


I've isolated an interesting cDNA from a human Clontech library that could
only have been made via transplicing from mRNAs originating from 2 different
chromosomes.  It seems to be real, but before I follow up on it I was
wondering if anyone with experience in cloning has found cases where similar
artifacts might have arisen in library construction, or of other possible
problems with commercially available libraries. Or perhaps have advice on
how I might tell if it's an artefact of library construction.  In this case
the putative exon boundaries don't seem to follow what's been published for
the genes in question, but with splice donor/acceptor sites being pretty
degenerate in humans I don't really know if this is a big red flag or
not--the whole cDNA encodes a rather interesting open reading frame.
 
Thanks, Chris
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