From owner-celegans@net.bio.net Thu Sep 11 23:00:00 1997 Path: biosci!rutgers!nntp.upenn.edu!dsinc!spool.mu.edu!uwm.edu!vixen.cso.uiuc.edu!news-peer.sprintlink.net!news.sprintlink.net!Sprint!news.maxwell.syr.edu!news.new-york.net!news.columbia.edu!konichiwa.cc.columbia.edu!wc47 From: Wen Chenhui Newsgroups: bionet.celegans Subject: YAC truncation method Date: Fri, 12 Sep 1997 14:07:49 -0400 Organization: Columbia University Lines: 33 Message-ID: Reply-To: Wen Chenhui NNTP-Posting-Host: konichiwa.cc.columbia.edu Mime-Version: 1.0 Content-Type: TEXT/PLAIN; charset=US-ASCII 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@columbia.edu . ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 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@columbia.edu ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 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 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ .