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Nucleic acid releasing agent and method and application for quickly releasing nucleic acid

A nucleic acid release agent and nucleic acid technology, applied in the field of molecular biology, can solve the problems of environmental and experimental operator hazards, restrictions on automatic promotion, cumbersome operation steps, etc., and achieve low reagent cost, strong scalability, and simple operation procedures Effect

Active Publication Date: 2017-12-15
HUNAN YEARTH BIOTECHNOLOGICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Nucleic acid extraction and purification generally undergoes liquid method-spin column method-magnetic bead method. Both liquid method and spin column method involve the use of toxic organic substances, which are harmful to the environment and experimental operators.
The magnetic bead method requires a specially prepared magnetic bead buffer and specifically modified magnetic beads. From the perspective of cost and operation, both of them have high cost and cumbersome operation steps. At the same time, it also limits the automation process and the promotion of applications.

Method used

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  • Nucleic acid releasing agent and method and application for quickly releasing nucleic acid
  • Nucleic acid releasing agent and method and application for quickly releasing nucleic acid
  • Nucleic acid releasing agent and method and application for quickly releasing nucleic acid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] The nucleic acid release agent in this example is composed of 100 mM Tris-HCl, 50 mM NaCl, 0.1 mM EDTA, 0.01% SDS, 0.04% LLS and 2% betaine.

[0024] The rapid release nucleic acid method of the present invention is used for TaqMan SNP detection of oral cavity, whole blood and dried blood spot samples, which involves the detection of TaqMan probes for single nucleotide polymorphisms of three genes EXOC1, CNTN4 and RNF146, specifically as follows Steps to proceed:

[0025] a) The oral scraper is directly eluted with the nucleic acid release agent, oscillated and mixed, and the oral mixed solution is obtained;

[0026] b) Take 50 μL whole blood sample and add it to a 1.5 mL centrifuge tube containing 150 μL ddH2O, centrifuge to discard the supernatant, then add 100 μL of the nucleic acid release agent to shake and resuspend;

[0027] c) Dried blood spot samples were punched with a 3mm hole puncher and put into a 1.5mL centrifuge tube. Add 500μL ddH2O to the centrifuge tu...

Embodiment 2

[0056] In this embodiment, the nucleic acid rapid release method is compared with the traditional spin column method, and the specific steps are as follows:

[0057] In this example, the same piece of dried blood spot sample was detected; 6 pieces were punched out with a 3mm punch, placed in 1.5mL centrifuge tubes respectively, and named as dried blood spot 1, dried blood spot 2, dried blood spot 3, dried blood spot Blood spot 4, dried blood spot 5, dried blood spot 6.

[0058] (1) Nucleic acid rapid release method

[0059] Take dried blood spot samples numbered 1-3, add 500 μL of ultrapure water to the centrifuge tube, vortex and mix well, let stand at room temperature for 5 minutes, centrifuge at 12000 rpm for 3 minutes, remove the supernatant, and then add the 200 μL nucleic acid release solution to the centrifuge tube Incubate the above sample tube with nucleic acid release agent in a metal bath at 80°C for 5 minutes; centrifuge at 12,000 rpm for 3 minutes, transfer the s...

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Abstract

The invention belongs to the technical field of molecular biology, in particular to a nucleic acid releasing agent and a method and application for quickly releasing nucleic acid. The nucleic acid releasing agent is prepared from 5-500 mM of Tris-HCl, 50-1000 mM of NaCl, 0.1-10 mM of EDTA, SDS with the mass to volume ratio being 0.01-2%, LLS with the mass to volume ratio being 0.04-3% and betaine with the mass to volume ratio being 0.01-2%. According to the method for quickly releasing nucleic acid, nucleic acid in serum, plasma, urine, saliva, oral swabs, dried blood stains, forensic medicine samples and environmental samples can be quickly released, and the released nucleic acid can be applied to downstream experiments such as detection, clone, sequence analysis and molecular hybridization.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and in particular relates to a nucleic acid releasing agent, a method for rapidly releasing nucleic acid and an application thereof. Background technique [0002] Currently, different types of genetic testing involve nucleic acid extraction and purification steps. Nucleic acid extraction and purification generally undergoes liquid method-spin column method-magnetic bead method. Both liquid method and spin column method involve the use of toxic organic substances, which are harmful to the environment and experimental operators. The magnetic bead method requires a specially prepared magnetic bead buffer and specifically modified magnetic beads. From the perspective of cost and operation, both of them have high cost and cumbersome operation steps. At the same time, it also limits the automation process and the promotion of applications. . Therefore, it is an urgent problem to develop a r...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
CPCC12N15/1003C12Q2527/127C12Q2523/113C12Q2527/125
Inventor 陆利沈欢徐根明潘艺赵谦
Owner HUNAN YEARTH BIOTECHNOLOGICAL CO LTD
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