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Nucleic acid extraction and amplification detection kit and detection method thereof

A detection kit and the technology of the kit are applied in the fields of biochemical equipment and methods, microbial determination/inspection, DNA preparation, etc., which can solve the problems of complex configuration work, experimental errors, and high costs, and enhance the effect of nucleic acid cleavage. , the effect of enhancing the degree of purification

Active Publication Date: 2019-02-22
NINGBO AJCORE BIOSCIENCES INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Obviously, the traditional method has many steps, high cost, large demand for samples and easy to cause cross-contamination, which is not conducive to the rapid extraction and purification of nucleic acid samples by customers
[0004] In addition, in the real-time fluorescent PCR technology that is most widely used in nucleic acid detection, most of the reagents are not ready-to-use, resulting in the common problems of complicated operation, laborious and high cost in PCR detection, and complex configuration work is required before use, which is easy error or even failure of the test
Experimental errors are also caused by different personnel operations; some reagents are unstable at room temperature; one-step rapid nucleic acid extraction cannot be achieved; PCR detection reagents need to be manually configured on site, multiple steps need to be manually marked, and the detection instruments at each stage are independent. Facilitates fully automated processing of large numbers of test samples

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Prepare 40ML of nucleic acid lysate according to the scheme of the present invention, including CuHCl 5.6g / 10ML, ethylenediaminetetraacetic acid 0.03g / 10ML, trishydroxymethylaminomethane 0.06g / 10ML, NaCl 0.01g / 10ML, polyethylene glycol Octyl phenyl ether 2% (volume ratio), surfactant can be sodium lauryl sulfate, surfactant 0.075% (volume ratio), 1% (volume ratio) phenol, isoamyl alcohol mixture, wherein The volume ratio of phenol and isoamyl alcohol is 20:1, and the pH value of the lysate is adjusted to 4.

Embodiment 2

[0025] Prepare 40ML of nucleic acid lysate according to the scheme of the present invention, including CuHCl 5.85g / 10ML, ethylenediaminetetraacetic acid 0.04g / 10ML, trishydroxymethylaminomethane 0.07g / 10ML, NaCl 0.015g / 10ML, polyethylene glycol Octyl phenyl ether 10% (volume ratio), the surfactant can be mixed with polyvinylpyrrolidone and sodium lauroyl acid acid, surfactant 0.1% (volume ratio), 2.5% (volume ratio) phenol, isoamyl alcohol Mixed solution, wherein the volume ratio of phenol and isoamyl alcohol is 25:1, and the pH value of the lysate is adjusted to 4.5.

Embodiment 3

[0027]Prepare 40ML of nucleic acid lysate according to the scheme of the present invention, including CuHCl 5.75g / 10ML, ethylenediaminetetraacetic acid 0.038g / 10ML, tris 0.062g / 10ML, NaCl 0.011g / 10ML, polyethylene glycol The octyl phenyl ether is 2.7% (volume ratio), the surfactant is cetylmethyltriamine bromide, the surfactant is 0.85% (volume ratio), and 2.5% (volume ratio) of phenol is added to the nucleic acid lysate 1. Isoamyl alcohol mixed solution, wherein the volume ratio of phenol and isoamyl alcohol is 22:1, and the pH value of the lysate is adjusted to 4.

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PUM

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Abstract

The invention provides a nucleic acid extraction and amplification detection kit. The nucleic acid extraction and amplification detection kit is composed of a lysate kit and a PCR (Polymerase Chain Reaction) pipe. Nucleic acid lysate is filled in the lysate kit, and after a to-be-detected sample is added into the nucleic acid lysate, viral nucleic acid is released by virtue of the effect of the nucleic acid lysate, and purification is realized. After the sample is added into the nucleic acid lysate, the obtained product can be directly added into the PCR pipe for realizing PCR detection. A freeze-dried powder PCR amplification system reagent which is directly used for fluorescent quantitative PCR measurement is arranged in the PCR pipe. The freeze-dried powder PCR amplification system reagent is a product prepared by taking a PCR buffer solution, DNA polymerase, a reaction specific primer, a probe and a freeze-drying protective additive as raw materials by virtue of a freeze drying process.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a nucleic acid extraction and amplification detection kit and a detection method thereof. Background technique [0002] Real-time fluorescence quantitative PCR technology has been widely used in many fields such as molecular diagnosis of genetic diseases, clinical testing, animal and plant import and export quarantine, food safety monitoring, soil microbial detection and paternity testing. Since blood, food, soil and other samples contain a large number of inhibitors, such as hemoglobin, hemethemin, lactoferrin, humic acid, etc., they will have obvious inhibitory effect on conventional Taq DNA polymerase. Therefore, nucleic acid must be isolated and extracted from these samples to be tested, and then used for PCR amplification. Nucleic acid extraction is the first step in nucleic acid detection and one of the key methods in molecular biology. Provide the basis for ...

Claims

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Application Information

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IPC IPC(8): C12N15/10C12Q1/686
CPCC12N15/1003C12Q1/686
Inventor 周杰锋王德明
Owner NINGBO AJCORE BIOSCIENCES INC
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