一种快速、经济提取外周凝血基因组DNA方法的建立  

Establishment of a Rapid Method for Extracting Human Genomic DNA from Peripheral Clotted Blood

作　　者：许丽娟[1] 马骁[1] 王洋阳[1] 王静[1] 潘晴[1] 刘梅[1] 

机构地区：[1]江苏省分子医学生物技术重点实验室南京师范大学生命科学学院,江苏南京210046

出　　处：《现代生物医学进展》2011年第20期3946-3950,共5页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(30700200);江苏省自然科学基金(青年科技创新人才启动项目;BK2007599)

摘　　要：目的:建立一种经济、快速且高质量提取人体外周凝血DNA的方法。方法:摸索最佳的匀浆条件,对外周凝血块进行匀浆,采用KI法对匀浆液进行基因组DNA的提取,通过凝胶电泳、单重PCR和多重PCR检测凝血基因组DNA的提取产量和质量,并分别与常规的凝血基因组DNA提取方法,即蛋白酶K消化法,以及提取抗凝血基因组DNA的KI法进行比较分析。结果:最佳的匀浆条件为:39000 rmp,15秒。在此条件下提取的基因组DNA完整性好,纯度和产量与蛋白酶K消化法提取凝血DNA和KI法提取抗凝血DNA的结果相比,没有统计学差异。单重PCR和多重PCR也获得了理想的扩增结果。结论:与常规的外周凝血提取方法相比(蛋白酶K消化法),本方法节省了时间和成本,能快速、经济、有效地提取外周凝血基因组DNA,可用于后续的科研和临床诊断需要,解决了部分科研机构血液基因组DNA的样本来源问题。Objective： To establish a rapid, economical method for extracting genomic DNA from peripheral clotted blood. Methods： Explore a optimum homogenate condition, homogenate the clotted blood, extracting the genomic DNA by KI method. Use agarose gel electrophoresis, single PCR and multiplex PCR detect the genomie DNA extraction yield and quality. And compared with the traditional extraction methods, Proteinase K digestion method and KI method which extract DNA from anti-coagulated blood. Results： The optimum homogenate condition is 39000 rmp and 15 seconds. The genomic DNA was extracted under this condition has good integrity. There is no significant difference of purity and yield between this method and Proteinase K digestion and KI method. Single PCR and multiplex PCR also obtained good amplification results. Conclusion： Compared with traditional extraction methods （Proteinase K digestion method）,this method can save time and cost,which can extract DNA rapidly, economical and effectively, and can be used in clinical and research analysis, and can solve the blood genomic DNA sources of some of the research institutions.

关 键 词：凝血基因组DNA 匀浆 KI法 蛋白酶K消化法 

分 类 号：R446[医药卫生—诊断学] Q75[医药卫生—临床医学]