A 3.5 genome equivalent multi access YAC library: construction, characterisation, screening and storage.
The construction of a yeast artificial chromosome (YAC) primary gridded library of 35,000 clones from human lymphoblastoid (48,XXXX) cell line DNA is described. The average YAC size is approximately 350kb representing a greater than 3.5 times coverage of the genome. The library is stored at -70 degrees 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.
ICI Diagnostics, Northwich, Cheshire, UK.
R
R
Anand
J H
JH
Riley
R
R
Butler
J C
JC
Smith
A F
AF
Markham
D002460:Cell Line; D002874:Chromosome Mapping; D015825:Chromosomes, Fungal; D003001:Cloning, Molecular; D015723:Gene Library; D006801:Humans; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000235:genetics; D014170:Transformation, Genetic