A LDR-PCR Approach for Multiplex Polymorphisms Genotyping of Severely Degraded DNA with Fragment Sizes <100 bp
A LDR-PCR Approach for Multiplex Polymorphisms Genotyping of Severely Degraded DNA with Fragment Sizes <100 bp
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DOI：
10.1111/j.1556-4029.2009.01166.x
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发表时间：
2009-11-01
影响因子：
1.6
通讯作者：
Xuan, Jin-Feng
中科院分区：
文献类型：
--
作者：
Zhang, Zhen;Wang, Bao-Jie;Xuan, Jin-Feng
减少扩增子大小已成为分析法医样品中典型的降解 DNA 的主要策略。然而，当前的小型短串联重复聚合酶链式反应 (PCR) 和小型测序测定中的扩增子大小仍然不适合分析严重降解的 DNA。在这项研究中，我们提出了一种将连接酶检测反应与 PCR 结合起来的多重分型方法，可用于识别严重片段化 DNA 样本中的单核苷酸多态性和小规模插入/缺失。该方法采用热稳定连接来区分等位基因，并采用随后的PCR来增强信号。在这项研究中，使用四个多态性位点来评估该技术区分人工降解 DNA 片段大小等位基因的能力
Reducing amplicon sizes has become a major strategy for analyzing degraded DNA typical of forensic samples. However, amplicon sizes in current mini-short tandem repeat-polymerase chain reaction (PCR) and mini-sequencing assays are still not suitable for analysis of severely degraded DNA. In this study, we present a multiplex typing method that couples ligase detection reaction with PCR that can be used to identify single nucleotide polymorphisms and small-scale insertion/deletions in a sample of severely fragmented DNA. This method adopts thermostable ligation for allele discrimination and subsequent PCR for signal enhancement. In this study, four polymorphic loci were used to assess the ability of this technique to discriminate alleles in an artificially degraded sample of DNA with fragment sizes