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Method for detecting single base mutation, and application thereof

A single-base mutation and purpose technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems of poor DNA purification effect, poor genetic detection effect, complicated extraction process, etc., to avoid abuse Drugs and adverse drug reactions, predicting disease risk, and effective effects

Inactive Publication Date: 2019-09-24
武汉百翼生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] A gene is the basic unit of heredity. A DNA or RNA sequence carrying genetic information transmits the genetic information to the next generation through replication and guides the synthesis of proteins to express the genetic information it carries. The DNA extraction process is complicated, and the DNA may contain a small amount of RNA or protein, which makes the DNA purification effect poor, and the genetic detection effect is poor

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] A method for detecting a single base mutation, comprising the steps of:

[0030] S1. DNA extraction: extract a small amount of animal or plant tissue, put it into a sterilized test tube, add a lysis solution to the test tube, and place the test tube in a high-speed centrifuge for centrifugation. The time is controlled at 5 minutes, and the lysis solution is specifically 0.1 mol / L sodium chloride solution, 10mol / L Tris-HCl solution and 1% SDS solution, the pH of the lysis solution is 8.0;

[0031] S2, DNA separation; specifically include adding phenol-chloroform solution, chloroform-isoamyl alcohol solution, ice absolute ethanol and 5mol / L sodium chloride solution to the test tube after completing operation S1, and place the test tube again Perform centrifugation on a high-speed centrifuge, and stop centrifugation when white flocculent precipitates appear in the solution in the test tube.

[0032] S3. Purification of DNA: After S2 is completed, add 60% ethanol solution ...

Embodiment 2

[0048] A method for detecting a single base mutation, comprising the steps of:

[0049] S1. DNA extraction: extract a small amount of animal or plant tissue, put it into a sterilized test tube, add a lysis solution to the test tube, and place the test tube in a high-speed centrifuge for centrifugation. The time is controlled at 8 minutes, and the lysis solution is specifically 0.1 mol / L sodium chloride solution, 10mol / L Tris-HCl solution and 1% SDS solution, the pH of the lysis solution is 8.0;

[0050] S2, DNA separation; specifically include adding phenol-chloroform solution, chloroform-isoamyl alcohol solution, ice absolute ethanol and 5mol / L sodium chloride solution to the test tube after completing operation S1, and place the test tube again Perform centrifugation on a high-speed centrifuge, and stop centrifugation when white flocculent precipitates appear in the solution in the test tube.

[0051] S3. Purification of DNA: After S2 is completed, add 65% ethanol solution ...

Embodiment 3

[0067] A method for detecting a single base mutation, comprising the steps of:

[0068] S1. DNA extraction: extract a small amount of animal or plant tissue, put it into a sterilized test tube, add a lysis solution to the test tube, and place the test tube in a high-speed centrifuge for centrifugation. The time is controlled at 10 minutes, and the lysis solution is specifically 0.1 mol / L sodium chloride solution, 10mol / L Tris-HCl solution and 1% SDS solution, the pH of the lysis solution is 8.0;

[0069] S2, DNA separation; specifically include adding phenol-chloroform solution, chloroform-isoamyl alcohol solution, ice absolute ethanol and 5mol / L sodium chloride solution to the test tube after completing operation S1, and place the test tube again Perform centrifugation on a high-speed centrifuge, and stop centrifugation when white flocculent precipitates appear in the solution in the test tube.

[0070] S3, DNA purification: After S2 is completed, add 70% ethanol solution an...

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PUM

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Abstract

The present invention relates to the technical field of gene detection and discloses a method for detecting a single base mutation. The method comprises DNA extraction, DNA separation, DNA purification, DNA analysis, NDA amplification and DNA detection. The DNA extraction is convenient, at the same time, purity of DNA can be effectively ensured, besides, a detection effect is obvious, the method can know whether disease-causing genes of familial diseases exist and predicts risks of diseases, the genetic diagnosis method is used, and the method greatly improves sensitivity, can obtain results in a short period of time, can correctly select drugs, and avoids drug abuse and drug adverse reactions; and according to results of genetic testing, a specific treatment scheme can be developed to scientifically guide use of drugs and avoids drug toxic and side effects.

Description

technical field [0001] The invention relates to the technical field of gene detection, in particular to a method and application for detecting single base mutation. Background technique [0002] A gene is the basic unit of heredity. A DNA or RNA sequence carrying genetic information transmits the genetic information to the next generation through replication and guides the synthesis of proteins to express the genetic information it carries. The DNA extraction process is complicated, and the DNA may contain a small amount of RNA or protein, which makes the DNA purification effect poor and the genetic detection effect poor. Contents of the invention [0003] The present invention proposes a method and application for detecting single base mutations to solve the problems raised in the above-mentioned background technology. [0004] The present invention proposes a method for detecting a single base mutation, comprising the following steps: [0005] S1. DNA extraction: extra...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6858C12Q1/6806C12Q1/6886C12Q1/6883
CPCC12Q1/6858C12Q1/6806C12Q1/6886C12Q1/6883C12Q2600/106C12Q2600/156C12Q2525/131C12Q2531/113C12Q2565/125C12Q2523/113C12Q2523/32C12Q2527/125
Inventor 杨国华李婷王晓欣王磊
Owner 武汉百翼生物科技有限公司