A whole genome amplification method to generate long fragments from low quantities of genomic DNA

被引：45

作者：

Kittler, R ^{[1
]}

Stoneking, M ^{[1
]}

Kayser, M ^{[1
]}

机构：

[1] Max Planck Inst Evolutionary Anthropol, Dept Evolutionary Genet, D-04103 Leipzig, Germany

来源：

ANALYTICAL BIOCHEMISTRY | 2002年 / 300卷 / 02期

关键词：

whole genome amplification; degenerate oligonucleotide-primed PCR; DOP-PCR; STR; short tandem repeats; microsatellites;

D O I：

10.1006/abio.2001.5460

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Several whole genome amplification strategies have been developed to preamplify the entire genome from minimal amounts of DNA for subsequent molecular genetic analysis. However, none of these techniques has proven to amplify long products from very low (nanogram or picogram) quantities of genomic DNA. Here we report a new whole genome amplification protocol using a degenerate primer (DOP-PCR) that generates products up to about 10 kb in length from less than 1 ng genomic template DNA. This new protocol (LL-DOP-PCR) allows in the subsequent PCR the specific amplification, with high fidelity, of DNA fragments that are more than 1 kb in length. LL-DOP-PCR provides significantly better coverage for microsatellites and unique sequences in comparison to a conventional DOP-PCR method. (C) 2001 Elsevier Science.

引用

页码：237 / 244

页数：8

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