Plant & Animal Genomes XI Conference Plant & Animal Genomes XI ConferenceJanuary 11-15, 2003
Town & Country Convention Center
San Diego, CA
Poster: Large Insert Libraries
Quantitative traits are often controlled by unknown genes that can be quite easily assigned on genetic maps but hard to characterize. The availability of a genome-wide molecular physical maps based on BAC contig assembly could largely accelerate positional cloning for identification of relevant genes. Meantime, the existence of overlapping BAC contigs is crucial for large-scale genome sequencing. In the frame of an international effort for the construction of a global physical map of swine genome, we started the production of BAC contigs using a high resolution DNA sequencing gel-based BAC fingerprinting approach. We used our 5 X swine BAC library representing about 105000 clones of 140 kb average size. This library has already been extensively screened for ESTs and genetic markers. DNA from each clone is isolated and digested with a mix of EcoR1, Hind3 and Hae3 restriction enzymes. Fingerprints are generated by incorporation of ddATP labeled with either R6G or R100 or TAMRA. Fingerprints from three distinct clones labeled with the three different dyes are multiplexed, run out on Megabace sequencer and sized using Genetic Profiler software. An average of 70 fragments per clone are recovered in the range of 65 to 720 nucleotides. About 10 % of BAC clones produced less than 8 bands possibly due to a high number of repeats. BAC assembling in contigs is in process using FPC software.