Container for containing quantified liquid drop PCR water-in-oil emulsion and use method of container

Abstract

The invention provides a container for containing quantified liquid drop PCR water-in-oil emulsion and a use method of the container. The use method of a body structure of the container comprises the following steps that (a) in a water-in-oil emulsion generation stage, an oily solution is added into the body structure in an initial state, at least a portion of aqueous droplets containing nucleic acid molecules dispersed in the oily solution is received to form a water-in-oil emulsion; (b) in an amplification stage, the body structure is transferred into an amplification module, and at least part of the aqueous liquid drops containing the nucleic acid molecules are amplified in the independent liquid drops; (c) in the tiling stage, after amplification is completed, the body structure is connected with a tiled chip, so that at least part of the emulsion in the body structure is transferred into the tiled chip. Based on the concept of independent design, the container needs to play different roles in different stages, so that the digital PCR container can be used in an existing macroscopic amplification similar structure, and the system is lower in cost and simpler and more convenient to design.

Description

technical field [0001] The present application relates to a droplet-type PCR consumable tube, in particular to a water-in-oil emulsion container for quantitative droplet PCR and its use method. Background technique [0002] Digital polymerase chain reaction (dPCR) is an improvement to the traditional PCR method, which can be used to directly quantify the original copy number of nucleic acid sequences. The development of this idea originated from the idea of ​​independently amplifying in droplets and then detecting the amplified products. The idea of ​​using droplets as carriers to amplify nucleic acid molecular chains was first seen in the British Medical Research Council, and later transferred to the British Research and Innovation Foundation in a series of patent applications. The patent application number US submitted on December 16, 1999 09 / 464122 U.S. application, which mainly protects the early prototype of the split droplet thinking; the patent application number US10...

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Problems solved by technology

[0004] The purpose of this application is to propose a water-in-oil emulsion container for quantitative droplet PCR and its use method in view of the deficiencies in the above-mentioned prior art, so as to solve the problem that the amplification of the existing droplet type digital PCR system is basically in the In the micro-scale space, or in the integrated chip, the sealing requirements introduced are high, and a series of problems such as the inability to realize large-scale multi-sample processing and high tolerance requirements for various components, etc.

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