Continuous volume gradient capillary digital PCR (polymerase chain reaction) device and method for applying same

A capillary and digital technology, applied in the field of digital PCR, can solve the problem of high cost and achieve the effect of cost saving, good application prospect and accurate detection method

Active Publication Date: 2019-02-19
WEST CHINA HOSPITAL SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] In order to solve the problems of high cost and the need for preliminary experiments in the prior art, the present invention provides a continuous volume gradient capillary digital PCR device

Method used

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  • Continuous volume gradient capillary digital PCR (polymerase chain reaction) device and method for applying same

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Experimental program
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Embodiment

[0029] Example device use test

[0030] In this embodiment, the artificially synthesized EGFR gene is used as a test template.

[0031] 1. Method

[0032] 1.1 Continuous Volume Gradient Capillary Preparation

[0033] Use a capillary with an inner diameter of 1mm to make a continuous volume gradient capillary. Under the grill of an alcohol lamp, pull the capillary evenly and forcefully until it breaks, and trim the tip of the capillary to expose the tip hole unobstructed. A capillary having a thick end inner diameter of 1 mm, a thin end inner diameter of 20 μm, and a length of 3 cm was obtained.

[0034] 1.2 Preparation of oil phase

[0035] First prepare oil with a density close to the density of water. In this example, castor oil (0.98g / 1) and paraffin oil are mixed evenly according to a volume ratio of 3:1 and cholesterol is added. The ratio is 100ml castor oil and excess cholesterol is added to dissolve it to saturation State, add 0.5% Span 80 or 0.1% tween 60 surfactan...

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Abstract

The invention provides a continuous volume gradient capillary digital PCR (polymerase chain reaction) device which comprises a programmable thermal cycler, a set of capillaries and optical signal acquisition equipment. The sizes of the cross sections of the capillaries continuously change. The continuous volume gradient capillary digital PCR device has the advantages that the cost can be saved, digital PCR operation steps can be simplified, and the continuous volume gradient capillary digital PCR device can be high in DNA (deoxyribonucleic acid) quantification accuracy.

Description

technical field [0001] The invention relates to the field of digital PCR, in particular to a continuous volume gradient capillary digital PCR device and a use method thereof. Background technique [0002] Modern biological research, especially medical research, often involves the need for quantitative analysis of nucleic acids. For example, the detection of certain free DNA and its content in blood can guide the clinical diagnosis of certain cancers and monitor the effect of cancer treatment. In the past, relative quantification was mainly performed by fluorescent quantitative PCR, that is, a transcript of a gene with stable expression was selected as a reference to judge the amount of the target nucleic acid. However, this method is susceptible to the interference of non-target nucleic acid molecules (background noise), and is only suitable for qualitative or low-precision detection, and cannot meet the detection of some target nucleic acid molecules with particularly rare...

Claims

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Application Information

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IPC IPC(8): C12M1/38C12M1/00
CPCC12Q1/6851B01L3/502784B01L2200/0626B01L2200/12C12Q2563/159C12Q2531/113
Inventor 李孝锦
Owner WEST CHINA HOSPITAL SICHUAN UNIV
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