摘要
目的：探讨不同温度焚烧的股骨中的DNA的提取方法。方法：常压下以100℃～500℃对30例成人股骨进行焚烧，然后分别采用有机法、Microcon100＋Qiagen试剂盒（QIAquick PCR Purification kit）纯化法、有机法＋Qiagen试剂盒纯化法。并用ND-1000光度计进行DNA浓度测定，用Sinofiler扩增试剂盒进行荧光标记和复合扩增。结果：骨骼样本在20℃～500℃各组用3种方法检测的DNA浓度均值分别为10．1～0．0、67．2～0．0、49．8～0．0ng／μl。有机法在20℃～500℃各组的位点检出率为38．3％、28．3％、16．7％、9．4％、0．0％、0．0％；纯压法为97．3％、75．7％、45．6％、33．1％、12．5％、1．2％；有机法＋Qiagen试剂盒纯化法为98．5％、81．9％、53．5％、36．3％、7．7％、0．4％。结论：有机法提取的骨骼DNA浓度及位点检出率较其他两种方法低；未烧骨骼用后两种方法提取其DNA都能获得较高的浓度和较好的分型，焚烧温度在100~C～300℃之间的骨骼用有机法＋Qiagen试剂盒纯化法提取其DNA较好；400℃-500℃焚烧的骨骼用Microeon100＋Qiagen试剂盒纯化法提取其DNA较好。
Objective： To explore the DNA extraction techniques from femur burned under different temperatures. Methods： Under temperatures ranging from 20℃ to 500℃ at ordinary atmospheric pressure, 30 adult femur samples were burned, thereafter, the DNA was extracted by using organic method, microcon 100 and subsequent purification with QIA quick PCR purification kit, organic method and purification with QIA quick PCR purification kit, respectively. Then, DNA density was determined by using DN-1000 spectrophotometer and following by amplified by means of PCR. Results： The average densities of DNA extracted from burned femur under 20℃- 500℃ by using three different techniques were 10. 1 - 0.0 ng/μl, 67.2 - 0. 0 ng/μl and 49. 8 - 0. 0 ng/μl. When the femur were burned under 20℃, 100℃, 200℃, 300℃,400℃ and 500℃, the detection rates of DNA loci were respectively 38.3%, 28. 3%, 16.7%, 9.4%, 0.0% and 0. 0% using organic method, 97.3% ,75.5% ,45.6% ,33.1% 12.50% and 1.2% using combination of Microcon 100 and QIA quick PCR purification kit, and 98.5 %, 81.9 %, 53.5 %, 36.3 %, 7.7 % and 0.4% using combination of organic method and QIA quick PCR Purification Kit. Conclusion ： Organic technique is inferior to later techniques in DNA density and loci detection. Combination of organic method and QIA quick PCR purification kit is recommended to extract DNA from the bones burned under 20℃ to 300℃, while the combination of Microcon 100 and QIA quick PCR purification kit is preferred to extract DNA from the bones burned under 400℃ to 500℃.
出处
《解剖学杂志》
CAS
CSCD
北大核心
2010年第1期119-121,共3页
Chinese Journal of Anatomy
基金
辽宁省教育厅科技基金（202173376）
关键词
烧骨
DNA提取
纯化
短串联重复序列
burned bone
DNA extraction
purification
short tandem repeat
