Plant & Animal Genome V Conference
Town & Country Hotel, San Diego, CA, January 12-16, 1997.
PAG-V: W12 - MOLECULAR CYTOGENEITC APPROACHES FOR THE GENERATION OF GENETIC MAPS
W12
MOLECULAR CYTOGENEITC APPROACHES FOR THE GENERATION OF GENETIC MAPS
PONCE DE LEON, F. ABEL
311 Paige Laboratory, Dept. of Veterinary and Animal Science, University of Massachusetts, Amherst, MA. 01003-6410
Chromosome-specific DNA libraries have been generated from microdissected chromosomes for several farm animal species. Chronmosome microdissection and microcloning have been done by micromanipulation of chromosomes and downscaling standard DNA cloning reactions to one nanoliter volumes. Our strategy is based on complete Sau 3A I digestion of chromosomal DNA and ligation of adaptor molecules to chromosomal fragments. The adaptors are in turn used as the priming sites for polymerase chain reaction amplification of chromosomal fragments. The amplified products have been used to generate chromosome painting probes and chromosome-specific libraries spanning 30%, 7% and 50% of the bovine, swine and chicken genomes, respectively. Some of these small insert libraries have been screened with (AC)12 oligos to identify clones containing premapped microsatellite sequences. The availability of these premapped microsatellite markers has accelerated the generation of high density genetic maps for the set of microdissected chromosomes. Also, chromosome painting probes have been used to trace chromosome rearrangements among some ungulate species by heterologous fluorescent in situ hybridization.