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High-sensitivity virus nucleic acid extraction kit

A viral nucleic acid and high-sensitivity technology, applied in the field of molecular biology, can solve problems such as poor extraction effect, low sensitivity, and high equipment requirements, and achieve the effects of avoiding false negative problems, small pollution risks, and reducing pollution

Pending Publication Date: 2022-07-29
BIOTEKE CORP (WUXI) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

CN102220310B provides a method for extracting and purifying human saliva DNA, but its components contain proteinase K, the cost is high, and it needs to be transported in ice packs, which increases the transportation cost and limits the application scenarios, and when the formula is pretreated, it needs Warm bath at 56°C for 30 minutes, requiring more equipment
CN110684764B provides a lysate, a nucleic acid extraction kit and a nucleic acid extraction method for nucleic acid extraction. Although proteinase K is not required and cracked at room temperature, the extraction sensitivity is low, and the extraction of low-concentration samples cannot be achieved.
Most of the nucleic acid extraction kits on the market can only achieve the extraction lower limit of 500copies / mL. In the face of lower concentration samples, the extraction effect is poor or even impossible, resulting in false negatives and misjudgment

Method used

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  • High-sensitivity virus nucleic acid extraction kit
  • High-sensitivity virus nucleic acid extraction kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] A high-sensitivity nucleic acid extraction kit, including the following components:

[0032] Virus lysate: 3M guanidine isothiocyanate, 3M sodium iodide, 50g / L SLS, 100mL / L Triton-100, 100mM EDTA-2Na, 200mL / L isopropanol;

[0033] Magnetic bead buffer: magnetic beads with a concentration of 50 mg / mL, proclin 300 at 0.1 mL / L;

[0034] Rinse solution 1: 1M sodium chloride, 600ml / L isopropanol;

[0035] Rinse solution 2: 0.1M sodium chloride, 800ml / L ethanol;

[0036] Eluent: 1ml / L of DEPC.

[0037] Prepare 3 armoured RNA respiratory virus throat swab mock samples, take 200 μL samples respectively, add them to the lysate, and perform nucleic acid extraction according to the following methods:

[0038] (1) Pretreatment: The biological sample containing viral nucleic acid is lysed with a virus lysate, and the volume ratio of the virus lysate to the biological sample is (0.5-2): 1 to obtain a crude lysate;

[0039] (2) Nucleic acid adsorption: the crude lysate obtained in...

Embodiment 2

[0049]Virus lysate: 1.5M guanidine isothiocyanate, 2.5M sodium iodide, 10g / L SLS, 10g / L Triton-100, 100mM EDTA-2Na, 400ml / L isopropyl Alcohol, pH 7.4;

[0050] Magnetic bead buffer: magnetic beads with a concentration of 50 mg / mL, proclin 300 at 0.1 mL / L;

[0051] Rinse solution 1: 1M sodium chloride, 600ml / L isopropanol;

[0052] Rinse solution 2: 0.1M sodium chloride, 800ml / L ethanol;

[0053] Eluent: 1ml / L of DEPC.

Embodiment 3

[0055] Virus lysate: 2M guanidine isothiocyanate, 2M sodium iodide, 10g / L SLS, 50g / L Triton-100, 10mM EDTA-2Na, 200ml / L isopropanol, pH7.4;

[0056] Magnetic bead buffer: magnetic beads with a concentration of 50 mg / mL, proclin 300 at 0.1 mL / L;

[0057] Rinse solution 1: 1M sodium chloride, 600ml / L isopropanol;

[0058] Rinse solution 2: 0.1M sodium chloride, 800ml / L ethanol;

[0059] Eluent: 1ml / L of DEPC.

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Abstract

The invention discloses a high-sensitivity virus nucleic acid extraction kit, and belongs to the technical field of molecular biology. The virus nucleic acid extraction kit comprises a virus lysis solution, a magnetic bead buffer solution, a rinsing solution 1, a rinsing solution 2 and an eluent, the formula of the virus lysis solution is as follows: guanidine isothiocyanate of 1-5 M, sodium iodide of 1-5 M, SLS of 10-100 g / L, triton-100 of 10-100 mL / L, EDTA-2Na of 10-200 mM and isopropanol of 200-500 mL / L, and the pH value of the virus lysis solution is 7.0-8.0. The virus nucleic acid extraction kit disclosed by the invention has the advantages of small dosage, capability of extracting at room temperature, high sensitivity, high purity of extracted nucleic acid, wide applicability and the like.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, in particular to a high-sensitivity viral nucleic acid extraction kit. Background technique [0002] In molecular biology experiments, nucleic acid extraction is the most basic and most important link. The yield, purity and integrity of nucleic acid extraction are directly related to the success or failure of downstream nucleic acid detection, biological research or other new product development. Magnetic bead method nucleic acid extraction method refers to the use of superparamagnetic silica nano-magnetic microbeads (hereinafter referred to as magnetic beads) as the carrier, the nucleic acid is adsorbed in a high-salt solution by magnetic beads, and after magnetic separation and rinsing of impurities, in a low-salt solution. A method for nucleic acid extraction based on the principle of nucleic acid desorption from the surface of magnetic beads. This method does not require centrifuga...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
CPCC12N15/1013C12N15/1003
Inventor 刘新鹏李云鹏周志图
Owner BIOTEKE CORP (WUXI) CO LTD
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