摘要
目的优化蛋白酶K的用量和消化时间,用于提取肋软骨DNA。方法 30份肋软骨样本各取大小为0.2cm×0.3cm×0.4cm的软骨块5份,分别编为A～E组。采用不同蛋白酶K用量和消化时间的Chelex100方法,提取5组各30份软骨块DNA,进行DNA定量、采用Sinofiler试剂盒扩增后进行电泳检测,采用非参数检验法比较检验成功率。结果 A～E组样本中采用加入2μL蛋白酶K并消化30min的D组方法成功率最高(96.7%),组间差异具有统计学意义。结论采用蛋白酶K-Chelex100方法,选择加入2μL蛋白酶K,消化30min可有效提高肋软骨DNA分型检验成功率。
Objective To establish an optimized DNA extraction method for costal cartilage by adjusting the volume of proteinase K and digestion time.Methods DNA was isolated from 30 costal cartilages（0.2cm×0.3cm×0.4cm） of each group（A～E） with different volume of proteinase K and sample digestion time.The extracted DNA was quantified,and then amplified with the Sinofiler kit followed by electrophorese.Statistical analysis was carried out for different extraction methods.Results Compared with other treatments,it showed the highest successful rate（96.7%） with 2μL proteinase K and 30 mins＇ digestion time at 56℃（group D）.Differences between different groups were significant.Conclusion Proteinase K DNA extraction protocol,with 2μL proteinase K and 30 mins＇ digestion time at 56℃,can isolate DNA from costal cartilage with higher efficiency.
出处
《中国法医学杂志》
CSCD
2012年第1期54-55,共2页
Chinese Journal of Forensic Medicine
关键词
法医物证学
蛋白酶K
肋软骨
forensic biological evidence
protinase K
costal cartilage
