腐殖酸高度污染样本的短串联重复序列分析的改进
Improvement of short tandem repeat analysis of samples highly contaminated by humic acid.
作者信息
Seo Seung Bum, Jin Hong Xuan, Lee Hye Young, Ge Jianye, King Jonathan L, Lyoo Sung Hee, Shin Dong Hoon, Lee Soong Deok
机构信息
Department of Forensic Medicine, College of Medicine, Seoul National University, Seoul 110-799, South Korea.
出版信息
J Forensic Leg Med. 2013 Oct;20(7):922-8. doi: 10.1016/j.jflm.2013.08.001. Epub 2013 Aug 28.
We investigated several methods for obtaining successful short tandem repeat (STR) results from high-humic acid (HA)-content samples. DNA purification efficiency was tested for QIAquick(®) PCR Purification, QIAamp(®) DNA Investigator and Prepfiler™ Forensic DNA Extraction kits. HA-removal capacity of Inhibitor Remover and InhibitEX(®) Tablet was tested. Experiments on overcoming HA effects on STR amplification were conducted using an AmpliTaq Gold(®) DNA Polymerase and a TaKaRa Ex Taq™ Hot Start Version (Ex Taq HS) with BSA addition. QIAquick kit was most efficient in HA removal and Ex Taq HS showed high resistance to HA. Increasing the amounts of Taq polymerases and BSA addition were shown to be efficient in overcoming PCR inhibition, but BSA addition was superior to the former method. Inhibitor Remover and InhibitEX(®) Tablet did not positively affect the STR results. This study will help achieve better STR results with high-HA-content samples.
我们研究了几种从高腐殖酸（HA）含量样本中获得成功短串联重复序列（STR）结果的方法。对QIAquick® PCR纯化试剂盒、QIAamp® DNA Investigator试剂盒和Prepfiler™法医DNA提取试剂盒的DNA纯化效率进行了测试。测试了抑制剂去除剂和InhibitEX®片剂的HA去除能力。使用AmpliTaq Gold® DNA聚合酶和添加了牛血清白蛋白（BSA）的TaKaRa Ex Taq™热启动版本（Ex Taq HS）进行了克服HA对STR扩增影响的实验。QIAquick试剂盒在去除HA方面效率最高，Ex Taq HS对HA表现出高抗性。增加Taq聚合酶的用量和添加BSA在克服PCR抑制方面被证明是有效的，但添加BSA优于前一种方法。抑制剂去除剂和InhibitEX®片剂对STR结果没有积极影响。本研究将有助于从高HA含量样本中获得更好的STR结果。
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