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Lysis solution of saliva nucleic acid extracted by paramagnetic particle method

A salivary nucleic acid and lysate technology, applied in the field of lysate for extracting salivary nucleic acid by magnetic bead method, can solve the problems of poor nucleic acid concentration and limit the efficient application of magnetic bead method extraction method, and achieve high detection efficiency, good effect and nucleic acid purity. high effect

Inactive Publication Date: 2017-03-15
广东国盛医学科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the existing magnetic bead extraction method is generally aimed at serum and plasma, and the nucleic acid concentration obtained when it is directly used for nucleic acid extraction of saliva is not good, which limits the efficient application of the magnetic bead extraction method in saliva nucleic acid extraction.

Method used

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  • Lysis solution of saliva nucleic acid extracted by paramagnetic particle method
  • Lysis solution of saliva nucleic acid extracted by paramagnetic particle method
  • Lysis solution of saliva nucleic acid extracted by paramagnetic particle method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1: Nucleic acid extraction and detection using saliva as a biological sample

[0022] 1. Saliva sample processing: The saliva collection tube is the saliva preservation tube of Guangdong Guosheng Medical Technology Co., Ltd. Take 12 saliva samples without pretreatment, mix them upside down several times, and then add them directly into the lysate system for lysate digestion.

[0023] 2. Reagents:

[0024] a. Prepare 12 parts of lysate according to the ratio of raw material components, as shown in the table below:

[0025]

[0026] b. Magnetic beads-binding solution: a mixed solution obtained by uniformly dispersing 20ul magnetic beads in 200ul isopropanol;

[0027] c. Washing solution A: composed of 50mM Tris-HCl, 3M potassium thiocyanate, 0.5% CTAB, 2% PVP and 40% ethanol;

[0028] d. Washing solution B: 70% ethanol.

[0029] 3. Use the above reagents to extract nucleic acid from 12 saliva samples by magnetic bead method, the steps are as follows:

[00...

Embodiment 2

[0034]Example 2: Nucleic acid extraction and detection using blood as a biological sample

[0035] 1. Blood sample processing: Blood was collected in blood preservation tubes of Guangdong Guosheng Medical Technology Co., Ltd. Collect 10 human blood samples, mix them evenly upside down, and then directly add to the lysate system for lysate digestion.

[0036] 2. Reagents:

[0037] a. Lysis solution: mixed with 100mM Tris-HCl, 5M lithium chloride, 3% sodium laurate, 5% Triton-X-100, 1% CTAB and 50mM aminotriacetic acid;

[0038] b. Magnetic beads-binding solution: the mixed solution obtained by uniformly dispersing 10ul magnetic beads in 100ul ethanol;

[0039] c. Washing solution A: composed of 100mM Tris-HCl, 5M lithium chloride, 1% CTAB, 0.2% PVP and 40% ethanol;

[0040] d. Washing solution B: 70% ethanol.

[0041] 3. Use the above reagents to extract 10 human blood samples by magnetic bead nucleic acid extraction, the steps are as follows:

[0042] Dispense 200ul of bl...

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Abstract

The invention discloses lysis solution of saliva nucleic acid extracted by a paramagnetic particle method. The lysis solution is characterized by comprising 10-150mM of Tris-HCl, 1-10M of chaotropic salt, 0.1-10% of negative ion surface active agents, 1-20% of nonionic surface active agents, 0.1-4% of CTAB (cetyl trimethyl ammonium bromide) and 1-100mM of chelating agents. The lysis solution extracted by the paramagnetic particle method sufficiently and effectively splits cells, can effectively remove special impurities such as mucopolysaccharide, mucoprotein and food residues and other conventional impurities such as protein, fat, carbohydrate and salt in saliva, so that the extraction effect of the nucleic acid is better, the purity of the nucleic acid is higher, more efficient late detection is ensured, and detection results are more accurate. The lysis solution is applicable to extraction of the nucleic acid of biological samples such as whole blood, cell cultures, bacterial cultures, swabs and tissues.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a lysate for extracting saliva nucleic acid by a magnetic bead method. Background technique [0002] Nucleic acid extraction methods include traditional phenol-chloroform method, salting out method, membrane spin column method, etc. Traditional nucleic acid extraction methods have their own advantages and disadvantages, but the extraction effect is not good. The current mainstream nucleic acid extraction methods include silica gel membrane adsorption column method, immunoaffinity method of anti-DNA monoclonal antibody, magnetic bead nucleic acid extraction method widely used in automation platforms, etc. [0003] Magnetic bead nucleic acid extraction refers to the use of superparamagnetic silica nano-magnetic microbeads (hereinafter referred to as magnetic beads) as the carrier, through which the magnetic beads adsorb nucleic acid in a high chaotropic salt solution and decompose nuc...

Claims

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Application Information

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IPC IPC(8): C12N15/10C12Q1/68C40B50/06
CPCC12N15/1013C12Q1/6806C40B50/06
Inventor 邓杏飞曾汉兵霍云龙
Owner 广东国盛医学科技有限公司
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