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使用多重置换扩增提高法医DNA样本的扩增成功率

Increasing amplification success of forensic DNA samples using multiple displacement amplification.

作者信息

Ballantyne Kaye N, van Oorschot Roland A H, Mitchell R John

机构信息

Biology Division, Victoria Police Forensic Services Centre, 31 Forensic Drive, Macleod, 3085, VIC, Australia,

出版信息

Forensic Sci Med Pathol. 2007 Sep;3(3):182-7. doi: 10.1007/s12024-007-0017-2. Epub 2007 Oct 2.

Abstract

Multiple displacement amplification (MDA) is capable of amplifying nanogram template amounts of DNA with high accuracy to generate micrograms of representative product. Although MDA is able to amplify small template amounts (<100 pg), increased levels of preferential amplification and allelic dropout are observed. The use of molecular crowders (polyethylene glycol 400) can decrease the amplification bias and increase amplification success. We describe the use of standard and crowded MDA on low-concentration casework samples originating from blood, semen, saliva, hair, and trace DNA. While standard MDA produced no significant increases in STR genotyping success, the use of crowded MDA yielded an average increase of 15% in the number of alleles, with a significant decrease in the amplification bias, resulting in clearer, more-complete profiles from forensically relevant samples.

摘要

多重置换扩增(MDA)能够以高精度扩增纳克级模板量的DNA,以产生微克级的代表性产物。尽管MDA能够扩增少量模板(<100 pg),但会观察到优先扩增水平增加和等位基因脱失现象。使用分子拥挤剂(聚乙二醇400)可以减少扩增偏差并提高扩增成功率。我们描述了在源自血液、精液、唾液、毛发和微量DNA的低浓度法医样本上使用标准MDA和拥挤MDA的情况。虽然标准MDA在STR基因分型成功率上没有显著提高,但使用拥挤MDA使等位基因数量平均增加了15%,扩增偏差显著降低,从而从法医相关样本中获得更清晰、更完整的图谱。

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