Cloning the differences between two complex genomes.
The analysis of the differences between two complex genomes holds promise for the discovery of infectious agents and probes useful for genetic studies. A system was developed in which subtractive and kinetic enrichment was used to purify restriction endonuclease fragments present in one population of DNA fragments but not in another. Application of this method to DNA populations of reduced complexity ("representations") resulted in the isolation of probes to viral genomes present as single copies in human DNA, and probes that detect polymorphisms between two individuals. In principle, this system, called representational difference analysis (RDA), may also be used for isolating probes linked to sites of genomic rearrangements, whether occurring spontaneously and resulting in genetic disorders or cancer, or programmed during differentiation and development.
Lisitsyn
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Cold Spring Harbor Laboratory, NY 11724.
Lisitsyn
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Wigler
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M
D000256:Adenoviridae / Q000235:genetics; D010582:Bacteriophage lambda / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000737:chemistry*; D015342:DNA Probes; D004279:DNA, Viral; D005260:Female; D017353:Gene Deletion; D030342:Genetic Diseases, Inborn / Q000235:genetics; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D009369:Neoplasms / Q000235:genetics; D009693:Nucleic Acid Hybridization / Q000379:methods; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D012689:Sequence Homology, Nucleic Acid
D015342:DNA Probes; D004279:DNA, Viral; D004247:DNA