A 3.5 genome equivalent multi access YAC library: construction, characterisation, screening and storage

摘要

The construction of a yeast artifical chromosome (YAC) primary gridded library of 35,000 clones from human lymphoblastoid (48,XXXX) cell line DNA is described. The average YAC size is ～350kb representing a 3.5 times coverage of the genme. The library is stored at ?70°C as gridded clones on nylon filters impregnated with 20% glycerol and as glycerol suspensions of individual clones in microtitre plates providing a prolonged multi-user potential. To date we have used 14 single copy probes to screen this library by colony hybridisation as well as PCR and have isolated between 1 and 5 YAC clones for every probe.