Modified DOP-PCR for improved STR typing of degraded DNA from human skeletal remains and bloodstains
Modified DOP-PCR for improved STR typing of degraded DNA from human skeletal remains and bloodstains
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DOI：
10.1016/j.legalmed.2015.10.013
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发表时间：
2016-01-01
期刊：
影响因子：
1.5
通讯作者：
Budowle, Bruce
中科院分区：
文献类型：
--
作者：
Ambers, Angie;Turnbough, Meredith;Budowle, Bruce
法医和古代DNA样本由于暴露在恶劣环境中以及时间的推移，往往受到损坏且数量有限。人们已经提出了几种策略来应对降解和低拷贝模板带来的挑战，其中包括一种基于聚合酶链反应（PCR）的全基因组扩增方法，称为简并寡核苷酸引物PCR（DOP - PCR）。本研究评估了原始DOP - PCR引物的四种改良版本的效果，这些改良版本保留了至少一部分5'端确定序列，并改变了3'端的碱基数。使用这四种改良引物中的每一种，从受环境破坏的血迹、当代人类骨骼遗骸、美国内战时期的骨头样本以及二战士兵的骨骼遗骸中获得的短串联重复序列（STR）图谱，都比之前描述的DOP - PCR方法和常规STR分型所获得的图谱有所改善。此外，改良的DOP - PCR程序允许使用更大体积的DNA提取物，减少了对样本进行浓缩的需求，从而减轻了抑制剂同时浓缩的影响。由爱思唯尔爱尔兰有限公司出版。
Forensic and ancient DNA samples often are damaged and in limited quantity as a result of exposure to harsh environments and the passage of time. Several strategies have been proposed to address the challenges posed by degraded and low copy templates, including a PCR based whole genome amplification method called degenerate oligonucleotide-primed PCR (DOP-PCR). This study assessed the efficacy of four modified versions of the original DOP-PCR primer that retain at least a portion of the 5' defined sequence and alter the number of bases on the 3' end. The use of each of the four modified primers resulted in improved STR profiles from environmentally-damaged bloodstains, contemporary human skeletal remains, American Civil War era bone samples, and skeletal remains of WWII soldiers over those obtained by previously described DOP-PCR methods and routine STR typing. Additionally, the modified DOP-PCR procedure allows for a larger volume of DNA extract to be used, reducing the need to concentrate the sample and thus mitigating the effects of concurrent concentration of inhibitors. Published by Elsevier Ireland Ltd.