title: An improved method to identify BAC clones using pooled overgos.
abstract: Hybridization using overgo probes is an established approach for screening arrayed bacterial artificial chromosome (BAC) libraries. We have improved the use of overgos by increasing the yield of positive clones using reduced levels of radioisotopes and enzyme. The strategy involves labeling with all four radiolabeled nucleotides in a hot pulse followed by a cold nucleotide chase and then extending the exposure time to compensate for reduced specific activity of the probes. The resulting cost savings and reduced human exposure to radiation make the use of highly pooled overgo probes a more attractive approach for screening of BAC libraries from organisms with large genomes.
authors.affiliation: Department of Botany and Plant Sciences, University of California, Riverside, CA 92521-0124, USA.
authors.forename: Kavitha
authors.initials: K
authors.lastname: Madishetty
authors.forename: Pascal
authors.initials: P
authors.lastname: Condamine
authors.forename: Jan T
authors.initials: JT
authors.lastname: Svensson
authors.forename: Edmundo
authors.initials: E
authors.lastname: Rodriguez
authors.forename: Timothy J
authors.initials: TJ
authors.lastname: Close
mesh_terms: D022202:Chromosomes, Artificial, Bacterial; D015698:Genomic Library; D001467:Hordeum / Q000235:genetics; D015345:Oligonucleotide Probes; D011868:Radioisotopes
chemical_list: D015345:Oligonucleotide Probes; D011868:Radioisotopes