数字PCR测定DNA含量及测量结果不确定度评定  

Quantification of DNA by Digital Polymerase Chain Reaction and Uncertainty Evaluation of Determination Result

作　　者：柳方方[1] 张玲[2] 王晶[2] 陈大舟[2] 臧超[2] 于笑波[3] 秦培勇[1] 

机构地区：[1]北京化工大学,北京100029 [2]中国计量科学研究院,北京100013 [3]中国计量学院,杭州310018

出　　处：《化学分析计量》2013年第1期18-22,共5页Chemical Analysis And Meterage

基　　金：国家"十一五"科技部支撑项目(2008BAK41B03);中国计量科学研究院基本业务费项目(AKY0915;AKY1219)

摘　　要：数字PCR是一项不依赖校准物的DNA绝对定量技术,将含有DNA模板的反应溶液分配到大量独立的微室中并且进行扩增反应,通过统计反应室中的阳性信号来定量DNA的拷贝数。采用数字PCR技术对噬菌体λDNA的含量进行测定,并对测定结果进行不确定度评定。结果显示,由数字PCR测得的λDNA的含量为(2.43±0.35)μg/管(k=2),该结果与同位素稀释质谱法测量结果相吻合。The digital PCR （dPCR） has the potential to afford absolute quantitation of DNA copies without reference calibrator. Copy number quantification by dPCR relied on positive single molecule statistics in partitions Since the PCR solution was distributed across a large number of partitions and followed amplification. λ DNA samples were determined by dPCR and the uncertanity of determination result was evaluated. Result showed that phage λ DNA quantified by dPCR was （2.5 3 ± 0.3 3 ） μ g/vial （k=-2）, which was consistent with that acquired from isotope dilution mass spectrometry.

关 键 词：数字PCR DNA含量 不确定度 

分 类 号：O652[理学—分析化学]