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包含德国DNA数据库系统的短扩增子多重q8验证。

Validation of the short amplicon multiplex q8 including the German DNA database systems.

作者信息

Müller Kathrin, Braunschweiger Günter, Klein Rachel, Miltner Erich, Wiegand Peter

机构信息

Institute of Legal Medicine, University Hospital Ulm, Ulm, Germany.

出版信息

J Forensic Sci. 2009 Jul;54(4):862-5. doi: 10.1111/j.1556-4029.2009.01066.x. Epub 2009 May 7.

Abstract

We have developed a concept to enable the analyzing of degraded stains with limited DNA template quantity. Therefore we have constructed a short tandem repeat (STR) multiplex including the German DNA database systems (Q8). The amplicon lengths are smaller than 280 bp. For the validation of Q8 over 50 degraded samples were investigated. Amplifications were performed with "low copy number" PCR, the number of PCR cycles was increased to 33 and the reaction volume was decreased to 12.5 microL. Compared with the MPX2 and Nonaplex kit, the average success rate was increased using the Q8 kit by approximately 20% and 30%, respectively. The efficiency of a sensitive STR multiplex with reduced amplicon lengths was confirmed in comparing the success rates of Q8 for typing degraded samples and samples with limited amount of DNA template while partial profiles were observed with the majority of the samples using commercially available kits.

摘要

我们已经开发出一种概念,能够对DNA模板量有限的降解污渍进行分析。因此,我们构建了一个包含德国DNA数据库系统(Q8)的短串联重复序列(STR)复合扩增体系。扩增子长度小于280 bp。为了验证Q8,对50多个降解样本进行了研究。采用“低拷贝数”PCR进行扩增,PCR循环数增加到33个,反应体积减少到l2.5微升。与MPX2和九重试剂盒相比,使用Q8试剂盒的平均成功率分别提高了约20%和30%。通过比较Q8对降解样本和DNA模板量有限的样本进行分型的成功率,证实了具有较短扩增子长度的灵敏STR复合扩增体系的有效性,而使用市售试剂盒时,大多数样本观察到部分图谱。

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