YAC truncation method

Wen Chenhui wc47 at columbia.edu
Fri Sep 12 13:07:49 EST 1997


In the course of my recent effort to clone the sup-17 gene, I have used a
method to make truncated derivatives from a YAC clone. After two steps of
trimming off both ends of the YAC insert, I ended up with a much smaller 
YAC(about 60kb) that covers only the cosmid gap. After showing that this
YAC still rescued sup-17, I eventually cloned sup-17 in a relatively short
time.
This method can be useful to anyone who has to work with YACs, either
because his or her favorite gene lies in a cosmid gap, or because the
overlapping cosmid clones are rearranged or hard to purify DNA from. I am
willing to share this method with anyone interested. The method is
specific to YACs that are pYAC4 backboned and hosted by yeast strain
AB1380, as are the YACs of worm genome.
Any request for the protocol and DNA constructs involved can be sent to
wc47 at columbia.edu  . 

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Chenhui Wen
Department of Biochemistry and Molecular Biophysics
Columbia University
College of Physicians and Surgeons
New York, NY 10032

Tel: (212) 305-6931
Fax: (212) 305-1721
email: wc47 at columbia.edu
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It begins to appear that everything one does to gain a livelihood or for
pleasure is fattening, immoral,illegal, or, even worse, oncogenic.
               ---Pathological Basis of Disease,  5th Edition(1995), PP254
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