Butler John M, Shen Yin, McCord Bruce R
Biotechnology Division, National Institute of Standards and Technology, Gaithersburg, MD 20899-8311, USA.
J Forensic Sci. 2003 Sep;48(5):1054-64.
New multiplex PCR sets of commonly used short tandem repeat (STR) markers have been developed to produce PCR products that are reduced in size when compared to standard commercial STR kits. The reduction in size of these amplicons can facilitate the examination and analysis of degraded DNA evidence by improving amplification efficiency. This "miniSTR" approach will permit current forensic practitioners to use STR markers and instrumentation already present in their laboratories and to generate genotyping data that is directly comparable to reference samples and searchable through the FBI's Combined DNA Index System (CODIS) databases. This paper discusses the development of these new primer sets and presents some initial results in the analysis of degraded and aged DNA samples. A method for removal of problematic fluorescent dye artifacts is also described. Comparison studies in over 100 samples have verified that these miniSTR primers can provide fully concordant results to commercial STR kits and can provide improved signal from degraded DNA specimens. These miniplex sets should prove valuable in the analysis of samples where allele dropout and reduced sensitivity of larger STR alleles occurs.
已经开发出了新的常用短串联重复序列（STR）标记多重PCR试剂盒，以产生与标准商业STR试剂盒相比尺寸更小的PCR产物。这些扩增子尺寸的减小可通过提高扩增效率来促进对降解DNA证据的检测和分析。这种“微型STR”方法将使当前的法医从业者能够使用其实验室中现有的STR标记和仪器，并生成与参考样本直接可比且可通过联邦调查局的联合DNA索引系统（CODIS）数据库进行搜索的基因分型数据。本文讨论了这些新引物试剂盒的开发，并展示了对降解和陈旧DNA样本分析的一些初步结果。还描述了一种去除有问题的荧光染料假象的方法。对100多个样本的比较研究证实，这些微型STR引物可提供与商业STR试剂盒完全一致的结果，并可从降解的DNA标本中提供更强的信号。这些微型多重试剂盒在分析出现等位基因脱失和较大STR等位基因灵敏度降低的样本时应具有重要价值。
