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Nucleic acid fast extraction kit and application method thereof

A kit and nucleic acid technology, applied in the field of molecular biology, can solve the problems that nucleic acid extraction is difficult to meet the requirements of large-scale and high-throughput experiments, and achieve the effect of easy automation, low price and stable properties

Inactive Publication Date: 2017-06-30
ZUNYI MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The present invention aims at the problem that nucleic acid extraction in the prior art is difficult to meet the requirements of large-scale and high-throughput experiments, and improves a simple and fast magnetic separation kit based on magnetic separation technology

Method used

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Embodiment 1: Taking the whole blood sample as an example:

[0024] Step 1: Take 100 μL of anticoagulated blood, add 10 μL of mixed enzyme working solution, and bathe in water at 56°C for 5 minutes.

[0025] Step 2: Add 750 μL of lysis and adsorption solution and appropriate amount of magnetic nanospheres and invert the test tube up and down 5-10 times to fully mix and then place at room temperature for 3 minutes.

[0026] Step 3: Separate the magnetic beads with a magnetic separation rack and discard the supernatant.

[0027] Step 4: Wash twice with 800 μL washing solution, separate the magnetic beads and discard the supernatant.

[0028] Step 5: Rinse twice with 1000 μL washing solution, separate the magnetic beads and discard the supernatant.

[0029] Step 6. Place at room temperature for 3 minutes, evaporate ethanol, add 100 μL of eluent and pipette the precipitate up and down to fully suspend the microspheres, then place at room temperature for 3 minutes to obtai...

Embodiment 2

[0034] The difference between Example 2 and Example 1 is that the concentration of proteinase K is 20 mg / mL, and the concentration of lysozyme is 20 mg / mL.

[0035] In the lysis adsorption solution, the concentration of Gu-Hcl is 4.4mol / L, the concentration of Tris-Hcl is 10mmol / L, the concentration of EDTA is 1mmol / L, the volume ratio of isopropanol is 1 / 4, SDS is 0.1%, Triton-100 is 1%, β-mercaptoethanol is 1%.

Embodiment 3

[0036] The difference between Example 3 and Example 1 is that the concentration of proteinase K is 40 mg / mL, and the concentration of lysozyme is 40 mg / mL.

[0037] In the lysis adsorption solution, the concentration of Gu-Hcl is 8mol / L, the concentration of Tris-Hcl is 30mmol / L, the concentration of EDTA is 10mmol / L, the volume ratio of isopropanol is 1 / 6, SDS is 0.5%, Triton -100 is 2%, β-mercaptoethanol is 2%.

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Abstract

The application discloses a nucleic acid fast extraction kit, which includes a mixed enzyme working solution, a cracking adsorption liquid, a detergent liquid, a rinsing liquid and an eluate liquid that are individually prepared. The mixed enzyme working solution includes protease K and lysozyme. The cracking adsorption liquid includes Gu-HCl, Tris-HCl, EDTA, isopropanol, SDS, Triton-100, beta-mercaptoethanol and sodium citrate. The detergent liquid includes LiCl, NaCl, MOPS and ethanol. The rinsing liquid is ethanol. The eluate liquid is EDTA and Tris-HCl. The kit is suitable for extraction of genomic DNA of whole blood, culture cell, tissues, saliva, G<-> bacteria and G<+> bacteria. The purity of extracted DNA is high, and the DNA is suitable for downstream digestion and PCR biological operation. The kit is stale in reagent property, can meet common laboratory demands and can overcome defects of toxicity, long time consumption and tedious operation of a traditional method.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and specifically relates to a rapid nucleic acid extraction kit and an extraction method for extracting genomic DNA from common clinical samples such as whole blood, cultured cells, tissues, saliva, G-bacteria, and G+ bacteria based on magnetic beads . Background technique [0002] Nucleic acid separation and purification technology is a basic technology in molecular biology experiments. Nucleic acid extraction is inseparable from the initial target gene extraction to PCR product purification or "gel recovery", as well as the extraction of plasmids in gene cloning. purification. Nucleic acid separation and purification technology originated in the 1950s. In the 1970s and 1980s, the traditional nucleic acid precipitation, dissolution, separation and purification method was widely used and promoted. Conventional nucleic acid extraction methods involve steps such as cell lysis, organic s...

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12N15/1013
Inventor 孙智勇许朋
Owner ZUNYI MEDICAL UNIVERSITY
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