Nucleic acid extraction kit and application method thereof

A kit and nucleic acid technology, applied in DNA preparation, recombinant DNA technology, etc., can solve the problems of affecting trace nucleic acid detection, not easy to discard, affecting molecular biological reactions, etc., and reduce the number and time of waste liquid washing. , Reduce the extraction time and the residue of waste liquid, the effect of reducing the extraction time

Active Publication Date: 2018-10-09
ZYBIO INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The first type of automation is applied in the transfer process of magnetic beads. There is a "hanging liquid" phenomenon on the magnetic beads, and a small amount of impurities will remain in the next reagent, especially in the eluent, which will affect the subsequent molecular biological reactions, especially Affects the detection of trace nucleic acids; the second type of automated application is not easy to discard when magnetic beads are used to remove waste liquid, which also causes residues to affect subsequent molecular biological reactions

Method used

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  • Nucleic acid extraction kit and application method thereof
  • Nucleic acid extraction kit and application method thereof
  • Nucleic acid extraction kit and application method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] Embodiment 1 The application of the magnetic bead method extraction method of the present invention to the quantitative detection of hepatitis B virus (HBV) nucleic acid on the semi-automatic nucleic acid extraction instrument

[0065] 1. Experimental materials

[0066] 1. Sample: HBV nucleic acid quantitative international standard NIBSC 10 / 264 (8.5E+05IU / mL, genotype A2), from the National Institute for Biological Standards and Control (NIBSC); negative human serum, from seracare company.

[0067] 2. Reagents:

[0068] 2.1 Lysis binding solution: fatty acid methyl ester ethoxylate sodium sulfonate 0.5%, guanidine isothiocyanate 1M, sodium acetate 1M, ethylphenyl polyethylene glycol 1%, proteinase K 10%, magnetic beads 3%, The pH value of the lysis binding solution is 7.0;

[0069] 2.2 Washing buffer: sodium iodide 1 mM, Tris hydrochloride 10 mM, ethanol 75%, the pH value of the washing buffer is 5.0;

[0070] 2.3 Silicone oil;

[0071] 2.4 Elution buffer: 1 mM et...

Embodiment 2

[0104] Embodiment 2 The application of the magnetic bead method extraction method of the present invention to the quantitative detection of hepatitis B virus (HBV) nucleic acid on the full-automatic nucleic acid extraction system

[0105] 1. Experimental materials

[0106] 1. Sample: consistent with the sample in Example 1;

[0107] 2. Reagents:

[0108] 2.1 Lysis binding solution: fatty acid methyl ester ethoxylate sodium sulfonate 2%, guanidine hydrochloride 5M, potassium acetate 5M, ethylphenyl polyethylene glycol 3%, proteinase K 20%, magnetic beads 5%. The pH value of the binding solution is 9.0;

[0109]2.2 Washing buffer: potassium iodide 10 mM, Tris hydrochloride 50 mM, ethanol 75%, the pH value of the washing buffer is 7.0;

[0110] 2.3 Silicone oil;

[0111] 2.4 Elution buffer: EDTA 5 mM, Tris hydrochloride 50 mM, the pH value of the elution buffer is 8.0.

[0112] 3. Instruments: Zhongyuan Biotechnology ZS9600 automatic nucleic acid extraction system, ABI 7500 ...

Embodiment 3

[0128] Embodiment 3 The application of the magnetic bead method extraction method of the present invention to the quantitative detection of hepatitis C virus (HCV) nucleic acid on the semi-automatic nucleic acid extraction instrument

[0129] 1. Experimental materials

[0130] 1. Sample: HCV nucleic acid quantitative international standard NIBSC 14 / 150 (1.0E+05IU / mL, genotype 1a), from the National Institute for Biological Standards and Control (NIBSC); negative human serum, from seracare company;

[0131] 2. Reagents:

[0132] 2.1 Lysis binding solution: fatty acid methyl ester ethoxylate sodium sulfonate 2%, guanidine hydrochloride 5M, potassium acetate 5M, ethylphenyl polyethylene glycol 3%, proteinase K 20%, magnetic beads 5%. The pH value of the binding solution is 8.0;

[0133] 2.2 Washing buffer: potassium iodide 10 mM, Tris hydrochloride 50 mM, ethanol 75%, the pH value of the washing buffer is 6.0;

[0134] 2.3 Silicone oil;

[0135] 2.3 Elution buffer: 1 mM ethy...

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Abstract

The invention provides a nucleic acid extraction kit, which comprises pyrolysis combining liquid, washing buffer liquid and elution buffer liquid, wherein the pyrolysis combining liquid is prepared from 0.5 to 2 percent of FAME (fatty acid methyl ester) ethoxylate sodium sulfonate and 3 to 5 percent of magnetic beads; less silicone oil is added into the washing buffer liquid; the whole nucleic acid extraction time is reduced; the waste liquid residue is reduced; nucleic acid extraction purification effect is improved; meanwhile, the precision and the sensitivity of nucleic acid detection are also improved.

Description

technical field [0001] The invention relates to the field of biotechnology, specifically, the invention relates to a method for extracting nucleic acid by magnetic beads. Background technique [0002] Nucleic acid is one of the most basic substances of life and widely exists in all animal and plant cells and microorganisms. According to different chemical compositions, nucleic acid can be divided into ribonucleic acid (RNA) and deoxyribonucleic acid (DNA), which play the role of storing and transmitting genetic information. [0003] With the popularization of genetic testing, personalized medicine, prenatal diagnosis, etc., high-throughput sequencing, fluorescent quantitative PCR, gene chips and other molecular biology technologies that use nucleic acid as the detection object are more and more widely used. One of the key links in nucleic acid detection is the extraction and purification of nucleic acids from various biological samples. The efficiency and purity of nucleic ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
CPCC12N15/1013
Inventor 陈威刘松林
Owner ZYBIO INC
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