数字聚合酶链反应
协变量
实验设计
核酸
R包
化学
生物系统
统计
计算机科学
数学
聚合酶链反应
生物化学
生物
基因
作者
Robert M. Dorazio,Margaret E. Hunter
出处
期刊:Analytical Chemistry
[American Chemical Society]
日期:2015-10-13
卷期号:87 (21): 10886-10893
被引量:23
标识
DOI:10.1021/acs.analchem.5b02429
摘要
Statistical methods for the analysis and design of experiments using digital PCR (dPCR) have received only limited attention and have been misused in many instances. To address this issue and to provide a more general approach to the analysis of dPCR data, we describe a class of statistical models for the analysis and design of experiments that require quantification of nucleic acids. These models are mathematically equivalent to generalized linear models of binomial responses that include a complementary, log–log link function and an offset that is dependent on the dPCR partition volume. These models are both versatile and easy to fit using conventional statistical software. Covariates can be used to specify different sources of variation in nucleic acid concentration, and a model’s parameters can be used to quantify the effects of these covariates. For purposes of illustration, we analyzed dPCR data from different types of experiments, including serial dilution, evaluation of copy number variation, and quantification of gene expression. We also showed how these models can be used to help design dPCR experiments, as in selection of sample sizes needed to achieve desired levels of precision in estimates of nucleic acid concentration or to detect differences in concentration among treatments with prescribed levels of statistical power.
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