Monitoring the efficacy of hybrid selection during positional cloning: the search for BRCA1.
Positional cloning often requires isolation of candidate genes from a large, genetically defined region. Hybrid selection (direct cDNA selection, solution hybrid capture) is a rapid, simple procedure that has been used to identify expressed sequence tags (ESTs) from cloned genomic DNA. We used hybrid selection to screen a 600-kb region that includes the BRCA1 gene. From a set of 931 sequenced clones, we obtained 118 nonoverlapping candidate ESTs from ovary and lymphocyte cDNA. We analyzed the results of our hybrid selection experiments with particular attention to the overall completeness, efficiency, and background noise of the experiment. We introduce simple parameters that serve as measures of important aspects of the hybrid selection process in the context of positional cloning.
Hattier
T
T
Myriad Genetics, Inc., Salt Lake City, Utah 84108, USA.
Bell
R
R
Shaffer
D
D
Stone
S
S
Phelps
R S
RS
Tavtigian
S V
SV
Skolnick
M H
MH
Shattuck-Eidens
D
D
Kamb
A
A
D019313:BRCA1 Protein / Q000235:genetics*; D001943:Breast Neoplasms / Q000235:genetics*; D002886:Chromosomes, Human, Pair 17; D003001:Cloning, Molecular; D018076:DNA, Complementary; D005260:Female; D006801:Humans; D009693:Nucleic Acid Hybridization; D012641:Selection, Genetic; D016324:Sequence Tagged Sites
D019313:BRCA1 Protein; D018076:DNA, Complementary