Hybridization selections, using immobilized driver DNA, to enrich for clones unique to a library.
We have developed a technique which allowed us to isolate sex-specific repeats of chickens (Gallus g. domesticus) from a genomic library originally containing one sex-specific repeat clone per 300 clones. Using this plus/minus selection method, we were able to enrich a sub-library in which the sex-specific repetitive clones made up over half of the population (170-fold increase in representation). This enrichment technique used hybridization kinetics to collect clones which are bound (plus selection) or not bound (minus selection) to their respective driver DNAs immobilized on nitrocellulose paper. Plus selections enriched for repetitive sequences and removed from the library most unique sequences as well as vectors containing no inserted sequences. These plus-selected sub-libraries were enriched for repetitive clones, yet still contained sequences representing as little as 10(-4) of the genome. Minus selection removed repetitive sequences shared between the driver and the library. When a plus-selected sub-library was minus selected, the doubly selected sub-library was enriched in repetitive sequences present in the library but not the minus driver.
Love
J
J
Institute of Animal Physiology and Genetics Research, Edinburgh Research Station Roslin, Midlothian, United Kingdom.
Deininger
P
P
D000818:Animals; D002645:Chickens; D003001:Cloning, Molecular; D004247:DNA / Q000032:analysis*; D005260:Female; D015723:Gene Library; D008297:Male; D009693:Nucleic Acid Hybridization; D012091:Repetitive Sequences, Nucleic Acid; D012727:Sex Characteristics
D004247:DNA