摘要
目的尝试建立一种检测混合斑中精子细胞的方法。方法使用显微操作法捕获精子细胞,全基因组扩增(多重置换扩增)精子细胞DNA。结果对10管精斑检材的全基因组扩增,获得了高产、保真的产物。使用50μL体系对20个精子细胞直接进行全基因组扩增,省去了对起始模板的纯化过程,DNA扩增倍数达30000倍以上,片段长度大多在15 kb以上,其STRs复合扩增分型结果有可参照性。结论显微操作法可以有效捕获精子细胞,排除干扰,多重置换扩增可以提供足够量的产物用于法医DNA分析,该方法具有可行性。
Objective To establish a method of preparing and analyzing DNA of sperm cells from sample mixtures. Methods Sperm cells were captured by micromanipulation method, DNA template was prepared from these sperm cells, and then amplified by multiple displacement amplification （MDA）. Results Highyield of highly integrated DNA was produced by MDA from very small amount of DNA template of 20 sperm cells. The DNA could be amplified at least 30000-fold, and the length of most the DNA segments were greater than 15kb, providing sufficient amount of short tandem repeat （STR） products for DNA genotyping. Conclusion The micromanipulation method is efficient to isolate sperm cells from sample mixtures and multiple displacement amplification can provide sufficient amount of DNA products for STR genotyping. The method seems to be feasible in detecting DNA from rape cases.
出处
《法医学杂志》
CAS
CSCD
2007年第4期286-289,共4页
Journal of Forensic Medicine
关键词
法医物证学
显微操作法
混合斑
精子细胞
多重置换扩增
全基因组扩增
短串联重复序列
forensic physical evidence
micromanipulation method
sample mixtures
sperm cells
multiple displacement amplification
genome amplification
STR
