Genomic Library Construction

ed beaty ed_beaty at qms1.life.uiuc.edu
Sun Dec 12 17:35:36 EST 1993


Hi!  I'm about to construct a genomic library to isolate a gene by colony
hybridization using a related gene as a probe, and also by genetic
complementation of an E. coli with a temp. sensitive copy of the same gene.
I have two questions:  How can I randomly cleave my genomic DNA into pieces
of about 8-12 kb?  I've heard of sonication, vortexing and passage through
a syringe needle to break the DNA, but I have no references.  Also, any
information on ligating the cleaved genomic vectors into plasmid vectors (I
was thinking of using pBluescript) would be really useful.

Also, I was trying to look on the University of Indiana Bionet archives for
this information, but the gopher system I'm using seems really slow.  It
shows me about a hundred files with such informative names as "RE>
libraries" and "RE>RE>RE>RE>libraries", but it takes about a full minute to
open up one of them.  Does anyone know of a faster way to get at the
archives (software-wise), or should I accept the fact that the digital
highway has a 10 mph speed limit sign on it?

As always, grateful for any assistance!

-Ed Beaty

Ed_Beaty at qms1.life.uiuc.edu
University of Illinois 
Department of Microbiology
"I'm not afraid of computers, but they're terrified of me..."



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