Construction Of A Bacterial Artificial Chromosome(BAC) Library Pool Of Chinese Fine-Wool Merino Sheep And The Control Of Quality

Bacterial artificial chromosome (BAC) libraries are the most convenient, reliable and widely used genomic libraries in genome research of organisms with highly complex genomes. In this study, a constructed sheep BAC library was copied. This library consists of about 190,000 clones arrayed in 497 384-well microtiter plates. Not I restriction analysis of 400 white clones picked randomly indicated the average insert size of BAC library is 133 kb.Theoretically,the probability to find any unique sequence in library screening is 99.93%. Tests on randomly selected clones demonstrated that they are very stable after serial growths . The process of duplicating is not contaminated through the analysis of PFGE.subsequently,construct a BAC pool through the method of pooling and superpooling, Not I restriction analysis of 400 BAC pooling clones picked randomly in BAC pools, indicated the average insert size is 133 kb ,which it is the same with the average insert size of BAC library,coming up to the demand of BAC pool.This pool provides a new resource for sheep genome analysis, including genome-wide physical mapping, genome sequencing and gene cloning and characterization, etc.