文献_DOP-PCR.pdfVIP

Proc. Natl. Acad. Sci. USA Vol. 93, pp. 14676–14679, December 1996 Genetics Whole genome amplification using a degenerate oligonucleotide primer allows hundreds of genotypes to be performed on less than one nanogram of genomic DNA VIVIAN G. CHEUNG AND STANLEY F. NELSON * Departments of Pediatrics and Biological Chemistry, University of California, Los Angeles, CA 90095 Communicated by Ronald W. Davis, Stanford University School of Medicine, Stanford, CA, September 30, 1996 (received for review May 2, 1996) ABSTRACT Genetic analysis of limiting quantities of We have investigated DOP-PCR amplification of total human genomic DNA play an important role in DNA forensics, genomic DNA at various starting amounts for yield and coverage paleoarcheology, genetic disease diagnosis, genetic linkage using microsatellite repeats. DOP-PCR was developed to allow analysis, and genetic diversity studies. We have tested the an unselected amplification of virtually any source DNA, has ability of degenerate oligonucleotide primed polymerase chain been specifically applied to cytogenetic techniques, and results in reaction (DOP-PCR) to amplify picogram quantities of hu- a more uniform signal than interspersed repeat-based methods of man genomic DNA for the purpose of increasing the amount whole genome amplification (7). In a single PCR tube several low of template for genotyping with microsatellite repeat markers. temperature annealings and extensions are performed to allow DNA was uniformly amplified at a large number of typable loci many binding sites in the human genome. After several low throughout the human genome with starting template DNAs temperature annealings and extensions, the annealing tempera- from as little as 15 pg to as