CAJ | 학술논문

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、Microcon 100+Qiagen试剂盒(QIAquick(R) 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℃～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Microcon 100+Qiagen试剂盒纯化法提取其DNA较好.
목적:탐토불동온도분소적고골중적DNA적제취방법.방법:상압하이100℃～500℃대30례성인고골진행분소,연후분별채용유궤법、Microcon 100+Qiagen시제합(QIAquick(R) PCR Purification kit)순화법、유궤법+Qiagen시제합순화법.병용ND-1000광도계진행DNA농도측정,용Sinofiler확증시제합진행형광표기화복합확증.결과:골격양본재20℃～500℃각조용3충방법검측적DNA농도균치분별위10.1～0.0、 67.2～0.0、 49.8～0.0ng/μl.유궤법재20℃～500℃각조적위점검출솔위38.3%、 28.3%、 16.7%、 9.4%、 0.0%、 0.0%;순압법위97.3%、 75.7%、 45.6%、 33.1%、 12.5%、 1.2%;유궤법+Qiagen시제합순화법위98.5%、 81.9%、 53.5%、 36.3%、 7.7%、 0.4%.결론:유궤법제취적골격DNA농도급위점검출솔교기타량충방법저;미소골격용후량충방법제취기DNA도능획득교고적농도화교호적분형,분소온도재100℃～300℃지간적골격용유궤법+Qiagen시제합순화법제취기DNA교호;400℃～500℃분소적골격용Microcon 100+Qiagen시제합순화법제취기DNA교호.
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 10.10.0ng/μl, 67.20.0ng/μl and 49.80.0ng/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% 12.5% 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℃.