A device for PCR quantification of samples with unknown nucleic acid concentration

Abstract

The invention provides a device for PCR quantification of unknown nucleic acid concentration samples, comprising a pressurizer (1), a plurality of closed containers (2), a plurality of connecting pipes (4) and an oil phase container (6), the closed container (2) For placing samples, one end of a plurality of connecting pipes (4) is converging and communicating, and the other end is respectively communicating with different airtight containers (2); the oil phase container (6) is communicating with the converging and communicating ends of the connecting pipes (4) ; The pressurizer (1) is used to inflate and pressurize the system formed by multiple airtight containers (2) and multiple connecting pipes (4); the volumes of the multiple airtight containers (2) are different, and the multiple connections The volumes of the flow channels of the tubes (4) are all equal, or the volumes of the multiple airtight containers (2) are all the same, while the volumes of the flow channels of the multiple connecting tubes (4) are not equal. The device can perform PCR quantification of samples with unknown nucleic acid concentration in a more simplified manner, which is beneficial to saving labor time and improving work efficiency.

Description

technical field [0001] The invention relates to the technical field of digital PCR quantification, in particular to a device for PCR quantification of samples with unknown nucleic acid concentration. Background technique [0002] Polymerase chain reaction (PCR) is a molecular biology technology used to amplify specific DNA fragments. Digital PCR is the third generation of PCR technology, known as the most advanced nucleic acid quantitative technology, mainly using microfluidics Methods Dilute the DNA or RNA solution in a large amount, and disperse it into the micro-droplets of the chip. The number of DNA templates in each droplet is less than or equal to 1. After PCR cycles, the nucleic acid is detected according to the number of droplets that generate fluorescent signals. Absolute Quantification of Molecules. [0003] According to statistical laws, in order to ensure the accuracy of digital PCR, it is often required that the ratio of the number of micro-droplets with fluor...

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Problems solved by technology

[0004] However, when quantifying a sample with an unknown nucleic acid concentration, since it is unknown how many nucleic acid molecules are contained in the sample, it is impossible to determine how many droplets need to be generated to ensure that the proportion of microdroplets with fluorescent signals to the total number of droplets meets the requirements. For this reason, it is necessary to conduct multiple tests on the same sample to find the experimental results that meet the requirements of the above ratio, and use the experimental results as the basis for calibration to determine the concentration of the nucleic acid solution.

[0005] At present, since it is necessary to repeat many identical test operations for the same sample for many times, it will take up a lot of time for the staff. Therefore, how to simplify the test operation to save labor time and improve work efficiency has become an important issue in this field. Technologists urgently need to solve technical problems

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