摘要
数字PCR是一项不依赖校准物的DNA绝对定量技术,通过统计(泊松分布)反应室中的阳性信号来定量DNA的拷贝数。该文对转基因油菜Ms8质粒分子进行构建和纯度分析后,采用荧光定量PCR对质粒分子的可替代性、均匀性和稳定性进行考察,采用数字PCR对质粒分子进行定值,并对测量结果进行不确定度评定。结果表明,数字PCR测得pMs8的外源基因和内源基因的比值及其不确定度为1.12±0.13,k=2。
The digital PCR( d PCR) has the potential to afford absolute quantitation of DNA copies without reference calibrator. Copy number quantificated by d PCR( Poisson distribution) relied on positive single molecule statistics. On this paper,after construction and purity of genetically modified rapeseed Ms8 of plasmid,q PCR was used to certify the homogeneity,stability and commutability,d PCR was used to certify the value. Also,the uncertanity of determination result was evaluated. The analysis results show that the ratio and uncertainty of exogenous gene and endogenous gene of pMs8 was 1. 12 ± 0. 13,k = 2.
出处
《中国测试》
北大核心
2017年第S1期15-20,共6页
China Measurement & Test
