Quantitative analysis method of digital PCRs of different volume

Abstract

The invention provides a quantitative analysis method of digital PCRs of different volume. Through the method, a corresponding standard deviation sigma and confidence interval of ln(c) can be obtained; the nucleic acid concentration c of a to-be-tested nucleic acid amplification reaction solution can be obtained through the corresponding standard deviation sigma and confidence interval of the ln(c), so that the initial copy number of DNA contained in the to-be-tested nucleic acid amplification reaction solution can also be known. The quantitative analysis method of the digital PCRs of different volumes can achieve the detection dynamic range of 5 orders of magnitudes by using less than 200 micro droplets, and enlarge the detection dynamic range of a digital PCR detection instrument. Moreover, the performance of the method is comparable to a single-volume digital PCR with 12,000 micro droplets, thereby reducing the instrument cost and reducing the consumable cost.

Description

technical field [0001] The invention relates to the field of digital PCR quantitative analysis, in particular to a method for quantitative analysis of digital PCR with different volumes. Background technique [0002] Digital PCR (Digital PCR, dPCR) is an absolute quantitative technique for nucleic acid molecules. Compared with qPCR, digital PCR allows you to directly count the number of DNA molecules, which is the absolute quantification of the starting sample. Quantitative PCR relies on a standard curve or a reference gene to determine the amount of nucleic acid, while digital PCR allows you to directly count the number of DNA molecules, which is the absolute quantification of the starting sample. However, the current quantitative detection of digital PCR, whether it is microwell or droplet digital PCR technology, adopts single-volume digital PCR technology. [0003] The upper limit of quantification of single-volume digital PCR mainly depends on the volume and number of ...

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Problems solved by technology

[0004] Based on this, it is necessary to provide a quantitative analysis method of different volumes of digital PCR with a large dynamic range for the problem of low sensitivity of the quantitative detection method of single volume digital PCR.

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