Identification and characterization of 23 RFLP loci by screening random cosmid genomic clones.
As part of our search for polymorphic DNA probes, we have screened cosmids from a human genomic DNA library for their ability to reveal RFLPs. A total of 101 randomly isolated cosmid clones were tested in Southern hybridizations for polymorphic band patterns. Fifty-four of these clones revealed RFLPs with one or more of nine restriction enzymes. Twenty-three of these clones have been further characterized and assigned to 10 different chromosomes by linkage analysis or by hybridization to panels of human-hamster hybrid cell lines. Fifteen of the probes have heterozygosities greater than or equal to .5. The relative efficiency of RsaI and PstI restriction enzymes in detecting polymorphism was different from results obtained with libraries constructed in bacteriophage vectors. Screening randomly selected cosmid probes is an efficient method for detecting RFLPs.
Bowden
D W
DW
Human Genetics Department, Collaborative Research, Incorporated, Waltham, MA 02154.
Müller-Kahle
H
H
Gravius
T C
TC
Helms
C
C
Watt-Morgan
D
D
Green
P
P
Donis-Keller
H
H
D000483:Alleles; D003360:Cosmids; D008040:Genetic Linkage; D006801:Humans; D010375:Pedigree; D011110:Polymorphism, Genetic; D012150:Polymorphism, Restriction Fragment Length