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Kit for nucleic acid extraction based on magnetic bead method and application thereof

A kit and magnetic bead method technology, which is applied in the field of nucleic acid extraction based on the magnetic bead method, can solve the problems of operator safety threats, long extraction process time-consuming, and non-negligible toxicity, so as to shorten the extraction time and reduce the time required for extraction and human labor, the effect of high degree of automation

Inactive Publication Date: 2019-03-22
GUANGDONG FOOD & DRUG VOCATIONAL COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these methods all have some disadvantages, such as the phenol-chloroform extraction method can obtain better quality nucleic acid, the toxicity of the chemical substances (such as: chloroform, phenol, etc.) used in it cannot be ignored, which has a great impact on the safety of operators. threat, and the entire extraction process takes a long time
In addition, the existing technology is not fully automated, most of them are semi-automatic, which brings great inconvenience to operators

Method used

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  • Kit for nucleic acid extraction based on magnetic bead method and application thereof
  • Kit for nucleic acid extraction based on magnetic bead method and application thereof
  • Kit for nucleic acid extraction based on magnetic bead method and application thereof

Examples

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Embodiment 1

[0050] This embodiment provides a kit for nucleic acid extraction based on the magnetic bead method, which includes a lysis binding solution, a digestion solution buffer, a washing solution I, a washing solution II and an eluent, and the lysis binding solution is composed of components with the following concentrations : Guanidine isothiocyanate 6mol / L, glycogen 80mg / L, Tris-Cl 20mmol / L, EDTA 2mol / L, Triton X-100 0.5% (v / v) and isopropanol 35% (v / v) , the solvent of the lysate-binding solution is sterile water;

[0051] Digestive fluid is made up of the component of following concentration: Nacl 10mmol / L and SDS 0.03% (w / v), the solvent of digestive fluid is; Buffer is the Triton X-100 sterile aqueous solution that concentration is 0.2% (v / v) The washing liquid I is made up of the following concentration components: sodium perchlorate 2mol / L and ethanol 50% (v / v), the solvent of the washing liquid I is sterile water; the washing liquid II is a concentration of 50% (v / v) v) et...

Embodiment 2

[0054] This embodiment provides a kit for nucleic acid extraction based on the magnetic bead method, which includes a lysis binding solution, a digestion solution buffer, a washing solution I, a washing solution II and an eluent, and the lysis binding solution is composed of components with the following concentrations : Guanidine isothiocyanate 5mol / L, glycogen 60mg / L, Tris-Cl 30mmol / L, EDTA 3mol / L, Triton X-100 0.2% (v / v) and isopropanol 30% (v / v) , the solvent of the lysate-binding solution is sterile water;

[0055] Digestive fluid is made up of the component of following concentration: Nacl 17mmol / L and SDS 0.01% (w / v), the solvent of digestive fluid is; The washing liquid I is made up of the following concentration components: sodium perchlorate 0.5mol / L and ethanol 40% (v / v), the solvent of the washing liquid I is sterile water; the washing liquid II is a concentration of 40% (v / v) ethanol sterile aqueous solution; the eluent is a Tris-Cl sterile aqueous solution with...

Embodiment 3

[0058] This embodiment provides a kit for nucleic acid extraction based on the magnetic bead method, which includes a lysis binding solution, a digestion solution buffer, a washing solution I, a washing solution II and an eluent, and the lysis binding solution is composed of components with the following concentrations : Guanidine isothiocyanate 8mol / L, glycogen 100mg / L, Tris-Cl 10mmol / L, EDTA 1mol / L, Triton X-100 0.4% (v / v) and isopropanol 40% (v / v) , the solvent of the lysate-binding solution is sterile water;

[0059] Digestive solution is made up of the components of following concentration: Nacl 20mmol / L and SDS 0.05% (w / v), the solvent of digestive solution is; The washing liquid I is made up of the following concentration components: sodium perchlorate 1.7mol / L and ethanol 60% (v / v), the solvent of the washing liquid I is sterile water; the washing liquid II is a concentration of 60% (v / v) ethanol sterile aqueous solution; the eluent is a Tris-Cl sterile aqueous solut...

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Abstract

The invention discloses a kit for nucleic acid extraction based on a magnetic bead method and application thereof and belongs to the field of molecular biotechnology. The kit comprises a cracking andbinding solution, a buffer solution, washing liquid I, washing liquid II and an eluent; the cracking and binding solution comprises the following components by concentration: 5-8 mol / L of isothiocyanic acid, 60-100 mg / L of glycogen, 10-30 mol / L of Tris-Cl, 1-3 mol / L of EDTA, 0.2-0.5 percent (v / v) of Triton X-100 and 30-40 percent (v / v) of isopropanol; a solvent of the cracking and binding solutionis sterile water. The cracking and binding solution in the kit disclosed by the invention integrates a cracking solution and a binding solution, so that the extraction efficiency is greatly enhanced;in addition, a reagent in the kit is pre-packed in an orifice plate, so that the automatic nucleic acid extraction by the magnetic bead method is realized.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and in particular relates to a kit and application for nucleic acid extraction based on a magnetic bead method. Background technique [0002] Nucleic acid extraction techniques mainly include phenol-chloroform extraction, alkali extraction and magnetic bead method. In the magnetic bead method, 96 deep-well plates are used with Smart32 automatic nucleic acid extractor for extraction. However, these methods all have some disadvantages, such as the phenol-chloroform extraction method can obtain better quality nucleic acid, the toxicity of the chemical substances (such as: chloroform, phenol, etc.) used in it cannot be ignored, which has a great impact on the safety of operators. threats, and the entire extraction process takes a long time. In addition, the prior art has not fully realized automation, and most of them are semi-automatic, which brings great inconvenience to operators. Con...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
CPCC12N15/1013
Inventor 宋卉迟海洋方春生王笑丹余永红
Owner GUANGDONG FOOD & DRUG VOCATIONAL COLLEGE
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