PCR (Polymerase Chain Reaction) experiment optimization method

An optimization method and experimental technology, which is applied in the field of PCR experiment optimization, can solve the problems that the measurement accuracy cannot be guaranteed, and cannot be predicted in advance, so as to achieve the effect of broadening the concentration detection range and improving the accuracy

Active Publication Date: 2018-03-09
领航医学科技(深圳)有限公司
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Problems solved by technology

[0003] Quantitative measurement of gene concentration by digital PCR based on Poisson distribution can have very high accuracy, but the measurement accuracy of the dilution in the dynamic range of unknown samples cannot be guaranteed or predicted in advance

Method used

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  • PCR (Polymerase Chain Reaction) experiment optimization method
  • PCR (Polymerase Chain Reaction) experiment optimization method
  • PCR (Polymerase Chain Reaction) experiment optimization method

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Embodiment Construction

[0032] In an exemplary embodiment, a method for optimizing a PCR experiment is provided. The method includes receiving the accuracy requirement for the experiment from the user, that is, the accuracy expectation value E, and determining an appropriate concentration calculation method according to the requirement, so as to optimize the PCR experiment.

[0033] Digital PCR based on the Poisson distribution can achieve extremely high precision in the quantitative measurement of genes. The present invention is aimed at large-sample PCR reaction units, which generally contain tens of thousands of reaction units. After the sample is fully diluted, the probability and quantity of single-molecule amplification reaction units will increase. The present invention establishes a mathematical model based on Poisson distribution by simulating numerical calculation MATLAB software to simulate the actual distribution of PCR target genes.

[0034] Such as figure 1 As shown, when the sample s...

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Abstract

The invention discloses a PCR (Polymerase Chain Reaction) experiment optimization method. The PCR experiment optimization method comprises the following steps: firstly, determining a low-order reaction unit negative rate PL and a high-order reaction unit negative rate PU according to a precision expected value E of PCR; if a reaction unit negative rate after PCR amplification ranges from the low-order reaction unit negative rate PL to the high-order reaction unit negative rate PU, calculating an initial concentration of a reactant by adopting a concentration calculation method based on a Poisson theory; otherwise, calculating the initial concentration of the reactant by adopting a real-time clustering concentration calculation method. According to the method disclosed by the invention, theprecision of calculating the initial concentration of the reactant in a PCR experiment is improved and a concentration detection range meeting precision requirements is expanded; meanwhile, the precision requirements also can be dynamically changed.

Description

technical field [0001] The invention relates to a PCR experiment optimization method. Background technique [0002] Digital PCR is to evenly distribute the fluorescent quantitative reaction system to a large number of tiny reaction units, and each tiny reaction unit does not contain or contain one or more target gene fragments. After the amplification is completed, the positive detection signal is generated for the fragment containing the target gene, and no detection signal is generated for the fragment not containing the target gene. The ratio of the number of positive reaction units judged by the terminal fluorescence signal to the total reaction units is calculated by statistical methods The copy number of the target gene in the original sample. [0003] Quantitative measurement of gene concentration by digital PCR based on Poisson distribution can have very high accuracy, but the measurement accuracy under dilution in the dynamic range of unknown samples cannot be guar...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G06F19/12G06F19/28C12Q1/6851
CPCC12Q1/6851G16B5/00G16B50/00C12Q2531/113C12Q2563/107C12Q2563/159
Inventor 程标
Owner 领航医学科技(深圳)有限公司
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