数字PCR测定DNA含量及测量结果不确定度评定.pdfVIP

22 1 Vol. 22No. 1 18 2013 1 CHEMICAL ANALYSIS AND METERAGE Jan. 2013 doi 10.3969j.issn.1008–6145.2013.01.005 PCR DNA * 1 2 2 2 2 3 1 , 1. 100029 2. 100013 3. 310018 PCR DNA DNA DNA PCR DNA PCR DNA k 2.430.35g ( =2) PCRDNA O652 A 1008–6145(2013)01–0017–05 Quantifi cation of DNA by Digital Polymerase Chain Reaction and Uncertainty Evaluation of Determination Result 1 2 2 2 2 3 1 Liu Fangfang , Zhang Ling , Wang Jing , Chen Dazhou , Zang Chao , Yu Xiaobo , Qin Peiyong (1. Beijing University of Chemical Technology Beijing100029, China 2. National Institute of Metrology Beijing100013, China 3. China Jiliang University Hangzhou310018 China) AbstratThe 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. DNA samples were determined by dPCR and the uncertanity of determination result was evaluated. Result showed that phage DNA quantified by dPCR was k (2.530.33) gvial ( =2)which was consistent with that acquired from isotope dilution mass spectrometry. Keywordsdigital PCR; DNA quantification; uncertainty PicoGreen4 DNA DNA DNA DNA 23 kb DNA 1985 1 Polymerase Chain Reaction PCR PCR 5–9 qPCRPCR 2–3DNA PCR PCR qPCR PCR DNA PCR (SYBR®green I) ( TaqMan®Probes) PCR