An anchored framework BAC map of mouse chromosome 11 assembled using multiplex oligonucleotide hybridization.
Despite abundant library resources for many organisms, physical mapping of these organisms has been seriously limited due to lack of efficient library screening techniques. We have developed a highly efficient strategy for large-scale screening of genomic libraries based on multiplex oligonucleotide hybridization on high-density genomic filters. We have applied this strategy to generate a bacterial artificial chromosome (BAC) anchored map of mouse chromosome 11. Using the MIT mouse SSLP data, 320 pairs of oligonucleotide probes were designed with an "overgo" computer program that selects new primer sequences that avoid the microsatellite repeat. BACs identified by these probes are automatically anchored to the chromosome. Ninety-two percent of the probes identified positive clones from a 5.9-fold coverage mouse BAC library with an average of 7 positive clones per marker. An average of 4.2 clones was confirmed for 204 markers by PCR. Our data show that a large number of clones can be efficiently isolated from a large genomic library using this strategy with minimal effort. This strategy will have wide application for large-scale mapping and sequencing of human and other large genomes.
Cai
W W
WW
Department of Molecular and Human Genetics, Howard Hughes Medical Institute, Baylor College of Medicine, One Baylor Plaza, Houston, Texas, 77030, USA.
Reneker
J
J
Chow
C W
CW
Vaishnav
M
M
Bradley
A
A
D000818:Animals; D002876:Chromosomes, Bacterial; D005819:Genetic Markers; D017403:In Situ Hybridization / Q000379:methods*; D051379:Mice; D008810:Mice, Inbred C57BL; D009841:Oligonucleotides / Q000235:genetics; D020161:Physical Chromosome Mapping / Q000379:methods*; D016133:Polymerase Chain Reaction; D012984:Software
D005819:Genetic Markers; D009841:Oligonucleotides