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Soil DNA extraction kit and soil DNA extraction method

A kit and soil technology, applied in the field of soil DNA extraction kits, can solve problems such as limiting the efficiency and quality of sample preparation, and tedious operation steps.

Inactive Publication Date: 2016-10-12
HUAIBEI NORMAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

Therefore, the above two extraction methods often involve a large number of toxic and harmful chemical reagents or organic solvents (such as phenol, chloroform, etc.) and tedious operation steps, which greatly limit the efficiency and quality of sample preparation.

Method used

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  • Soil DNA extraction kit and soil DNA extraction method
  • Soil DNA extraction kit and soil DNA extraction method
  • Soil DNA extraction kit and soil DNA extraction method

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Embodiment Construction

[0032] 1. The soil DNA extraction kit in the present embodiment is composed of the following six solutions that do not contain harmful organic solvents:

[0033] Solution A: a mixed solution of Tris, EDTA, SDS and NaCl, the concentration of Tris in the mixed solution is 50mM, the concentration of EDTA is 100mM, the mass concentration of SDS is 2%, the concentration of NaCl is 20mM, adjust the concentration of solution A with hydrochloric acid The pH value is 6.4;

[0034] Solution B: 4M KCl solution;

[0035] Solution C: a mixed solution of Tris and GuHCl, the concentration of Tris in the mixed solution is 10mM, the concentration of GuHCl is 6M, and the pH value of solution C is adjusted to 6.7 with hydrochloric acid;

[0036] Solution D1: a mixed solution of Tris, EDTA and GuHCl, the concentration of Tris in the mixed solution is 50mM, the concentration of EDTA is 10mM, the concentration of GuHCl is 3M, and the pH value of solution D1 is adjusted to 7.7 with hydrochloric aci...

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Abstract

The invention discloses a soil DNA extraction kit and a soil DNA extraction method, wherein the soil DNA extraction kit is composed of the following six solutions: solution A: a mixed solution of Tris, EDTA, SDS and NaCl and with pH value of 6.4; solution B: 4M of KCl solution; solution C: mixed solution of Tris and GuHCl and with pH value of 6.7; solution D1: a mixed solution of Tris, EDTA and GuHCl and with pH value of 7.7; solution D2: mixed solution of Tris and ethanol and with pH value of 6.7; and solution E: 10mM of Tris-HCl buffer solution and with pH value of 8.0-8.5. The kit can extract high-quality DNA suitable for technical operation in modern molecular biology from soil easily and quickly, and the method has the advantages of simplicity, quick speed, economy, high quality and the like.

Description

1. Technical field [0001] The invention relates to a kit for extracting soil DNA and a method for extracting soil DNA. 2. Background technology [0002] Modern molecular biology technologies, such as metagenomic sequencing technology, restriction fragment polymorphism (RFLP) technology, polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) technology, etc., are to further understand and use microorganisms to serve human beings important means are provided. However, DNA extraction and purification are the first and crucial steps in the application of these technologies. [0003] The current soil DNA extraction methods mainly include in situ lysis and cell extraction before lysis. Due to the complexity and heterogeneity of the soil itself, humic acid, clay minerals and other ions in the soil will affect the extraction and purity of soil DNA to varying degrees. Therefore, the above two extraction methods often involve a large number of toxic and harmfu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
CPCC12N15/1006C12Q2523/308
Inventor 熊明华李文波朱继荣王光利
Owner HUAIBEI NORMAL UNIVERSITY
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