Whole genome amplification using a degenerate oligonucleotide primer allows hundreds of genotypes to be performed on less than one nanogram of genomic DNA

被引:211
作者
Cheung, VG
Nelson, SF
机构
[1] UNIV CALIF LOS ANGELES,DEPT PEDIAT,LOS ANGELES,CA 90095
[2] UNIV CALIF LOS ANGELES,DEPT BIOL CHEM,LOS ANGELES,CA 90095
关键词
D O I
10.1073/pnas.93.25.14676
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Genetic analysis of limiting quantities of genomic DNA play an important role in DNA forensics, paleoarcheology, genetic disease diagnosis, genetic linkage analysis, and genetic diversity studies, We have tested the ability of degenerate oligonucleotide primed polymerase chain reaction (DOP-PCR) to amplify picogram quantities of human genomic DNA for the purpose of increasing the amount of template for genotyping with microsatellite repeat markers, DNA was uniformly amplified at a large number of template loci throughout the human genome with starting template DNAs from as little as 15 pg to as much as 400 ng. A much greater-fold enrichment was seen for the smaller genomic DOP-PCRs, All markers tested were amplified from starting genomic DNAs in the range of 0.6-40 ng with amplifications of 200- to 600-fold, The DOP-PCR-amplified genomic DNA was an excellent and reliable template for genotyping with microsatellites, which give distinct bands with no increase in stutter artifact on di-, tri-, and tetranucleotide repeats. There appears to be equal amplification of genomic DNA from 55 of 55 tested discrete microsatellites implying near complete coverage of the human genome, Thus, DOP-PCR appears to allow unbiased. hundreds-fold whole genome amplification of human genomic DNA for genotypic analysis.
引用
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页码:14676 / 14679
页数:4
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