A whole genome amplification method to generate long fragments from low quantities of genomic DNA
A whole genome amplification method to generate long fragments from low quantities of genomic DNA
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DOI：
10.1006/abio.2001.5460
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发表时间：
2002-01-15
影响因子：
2.9
通讯作者：
Kayser, M
中科院分区：
文献类型：
--
作者：
Kittler, R;Stoneking, M;Kayser, M
已经开发了几种全基因组扩增策略以从最少量的DNA预扩增整个基因组用于随后的分子遗传分析。然而，这些技术中没有一种被证明可以从非常低（纳克或皮克）量的基因组DNA扩增长产物。在这里，我们报告了一种使用简并引物（DOP-PCR）的新全基因组扩增方案，该方案从不到1 ng的基因组模板DNA中产生长度达约10 kb的产物。这种新的方案（LL-DOP-PCR）允许在随后的PCR中特异性扩增，具有高保真度，长度超过1 kb的DNA片段。与常规DOP-PCR方法相比，LL-DOP-PCR对微卫星和独特序列提供了显著更好的覆盖。(C)2001年，爱思唯尔科学。
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.