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Nucleic Acids Research, 1990, Vol. 18, No. 8 1951-1956
© 1990

GENOME STRUCTURE AND MAPPING

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

R. Anand, J.H. Riley, R. Butler, J.C. Smith and A.F. Markham

ICI Diagnostics Gadbrook Park, Northwich, Cheshire CW9 7RA, UK

Received February 16, 1990. Revised March 20, 1990. Accepted March 20, 1990.

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.


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