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使用Phi29 DNA聚合酶进行全基因组扩增,以实现对低DNA产量样本的遗传或基因组分析。

Whole genome amplification with Phi29 DNA polymerase to enable genetic or genomic analysis of samples of low DNA yield.

作者信息

Silander Kaisa, Saarela Janna

机构信息

National Public Health Institute, Helsinki, Finland.

出版信息

Methods Mol Biol. 2008;439:1-18. doi: 10.1007/978-1-59745-188-8_1.

Abstract

In many large genetic studies, the amount of available DNA can be one of the criteria for selecting samples for study. In the case of large population cohorts, selecting samples based on their DNA yield can lead to biased sample selection. In addition, many valuable clinical and research sample collections exist in which the amount of DNA is very small. Unbiased whole genome amplification (WGA) of such unique samples enables genomewide scale genetic studies that would have been impossible otherwise. Multiply primed rolling circle amplification (MPRCA) and multiple displacement amplification (MDA) methods are based on the same principle. The DNA amplification is non-PCR based and uses Phi29 DNA polymerase and random hexamer primers for unbiased whole genome amplification. MDA is used for linear DNA molecules, such as genomic DNA. This chapter reviews the various applications in which whole genome amplified DNA can be used, the types of commercial kits available, and the quality control steps necessary before using the DNA in the genetic studies.

摘要

在许多大型基因研究中,可用DNA的量可能是选择研究样本的标准之一。对于大型人群队列而言,基于DNA产量选择样本可能导致样本选择偏差。此外,存在许多有价值的临床和研究样本库,其中DNA的量非常少。对这类独特样本进行无偏差全基因组扩增(WGA),能够开展全基因组规模的基因研究,否则这是不可能实现的。多重引物滚环扩增(MPRCA)和多重置换扩增(MDA)方法基于相同的原理。DNA扩增不基于PCR,而是使用Phi29 DNA聚合酶和随机六聚体引物进行无偏差全基因组扩增。MDA用于线性DNA分子,如基因组DNA。本章综述了全基因组扩增DNA可用于的各种应用、可用的商业试剂盒类型,以及在基因研究中使用该DNA之前所需的质量控制步骤。

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