Gene mapping from a bovine 1;29 DNA library prepared with chromosome microdissection

✍ Scribed by S. M. Schmutz; T. G. Berryere; J. S. Moker; T. D. Thue; D. C. Winkelman

• Book ID: 104735744
• Publisher: Springer-Verlag
• Year: 1994
• Tongue: English
• Weight: 772 KB
• Volume: 5
• Category: Article
• ISSN: 0938-8990
• DOI: 10.1007/bf00352343

✦ Synopsis

Bovine gene mapping is progressing rapidly using syntenic group mapping based on somatic cell hybrids and linkage, and to a lesser extent on in situ hybridization. Single chromosome DNA libraries are a logical next step, and this was, therefore, the aim of our laboratory. Since we have access to several cattle with t(1 ;29) and this chromosome is readily distinguishable, we chose this as our first target--recognizing that we would not produce a "single" chromosome library in the strict sense because two autosomes are represented. We utilized an inverted microscope and a micromanipulator fitted with glass instruments pulled specifically to dissect off approximately 100 t(1 ;29) chromosomes per microdrop. A glass chamber made to accommodate a hanging drop was used to extract the DNA under a dissecting microscope. The DNA was then Cleaved with EcoRI and inserted in )~gtwes arms. Host cells were then infected with these phage and positive clones obtained. The first clone, isolated from this library by hybridization with a human collagen 6A1 cDNA, was mapped by in situ hybridization to bovine Chromosome (Chr) lq12-q14, near the centromere. The second clone, an anonymous DNA fragment (D1S11), was mapped to lq43-q46, near the terminal end.