PCR产物纯化增强STR分型强度的研究

目的研究PCR产物纯化对微量DNA的STR分型的影响。方法使用25pg/μL的9947标准基因组DNA为模板,标准程序扩增和STR分型检测。设立对照组A和实验组B、C、D。实验组分别使用分子筛(Qiagen DyeEX2.0spin kit)、超滤膜(Amicon Ultra-0.5,100KD)、亲和层析(Qiagen MinElute Column)3种DNA纯化方法,对照组不做任何处理。结果与对照组相比,3种方法均可显著提高STR分型强度(P峰高<0.001),平均峰高约为对照组的4倍,并且3种方法对提高STR分型强度无显著差异(P峰高=0.249)。结论 PCR产物纯化能显著提高微量DNA的STR分型强度,可用于骨骼、脱落细胞等微量DNA检材的检验。

Enhancement of STR profiles peak intensity using post-PCR purification

Objective This paper investigated the enhancement of STR profiles peak intensity using post-PCR purification, as it becomes increasingly important to gain DNA profile information from exceedingly trace levels of DNA. Methods The 9947 genome of 25pg/μl concentration was used as DNA template for STR profiling. The PCR product was divided into control group and three examined groups. The control group was electrophoresed as standard procedure, and in examined groups, PCR products were purified using molecule sieve method （Qiagen DyeEX2.0 spin kit）, ultrafiltration method （Amicon Uhra-0.5,100KD） and af finity chromatography method （Qiagen MinElute Column） separately before electrophoresis. Results In three examined groups, an approximately 4-fold increase in STR peak height （RFU） was observed relative to the standard conditions, while there is no sig- nificant peak height difference between three groups. Conclusions Post-PCR purification is an alternative tool for enhancing STR peak height and would be used in forensic casework.