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对非侵入性获取的猕猴精液DNA进行多次置换扩增后,序列质量得以保持。

Sequence quality is maintained after multiple displacement amplification of non-invasively obtained macaque semen DNA.

作者信息

Andrés Olga, Rönn Ann-Charlotte, Ferrando Aïnhoa, Bosch Montserrat, Domingo-Roura Xavier

机构信息

Genètica de la Conservació, Institut de Recerca i Tecnologia Agroalimentàries, Cabrils, Spain.

出版信息

Biotechnol J. 2006 Apr;1(4):466-9. doi: 10.1002/biot.200600011.

Abstract

Whole genome amplification protocols are revolutionizing the fields of molecular and conservation biology as they open the possibility of obtaining a large number of copies of a complete genome from minute amounts of sample. Multiple displacement amplification (MDA) is a whole genome amplification technique based on the properties of the phi29 DNA polymerase, which leads to a uniform representation of the genome with very low error rates. In this study we performed MDA on 28 macaque DNA samples extracted from blood or non-invasively collected semen from which we obtained mitochondrial control region sequences both before and after MDA. The length of the readable sequences was longer for the original samples than for the MDA products, but the number of unresolved positions was comparable both before and after MDA. We conclude that the MDA technique is useful for increasing the amount of DNA for sequencing mitochondrial regions in the case of non-invasively collected semen samples.

摘要

全基因组扩增方案正在革新分子生物学和保护生物学领域,因为它们开启了从微量样本中获得大量完整基因组拷贝的可能性。多重置换扩增(MDA)是一种基于phi29 DNA聚合酶特性的全基因组扩增技术,该技术能够以极低的错误率实现基因组的均匀代表性。在本研究中,我们对从血液中提取的28份猕猴DNA样本或非侵入性采集的精液进行了MDA,在MDA前后均获得了线粒体控制区序列。原始样本的可读序列长度比MDA产物的长,但MDA前后未解析位置的数量相当。我们得出结论,在非侵入性采集精液样本的情况下,MDA技术对于增加用于线粒体区域测序的DNA量是有用的。

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