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Method for quickly extracting DNA (deoxyribonucleic acid)

A fast, lysing solution technology, applied in the biological field, can solve the problems of complex operation, high cost, unable to meet the test requirements, etc., and achieve the effect of reducing the cost of use and simple steps.

Inactive Publication Date: 2012-09-19
亚能生物技术(深圳)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0013] Existing technical methods are too expensive and complicated to operate, and cannot meet the test requirements for testing a large number of samples, especially in clinical applications

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

[0047] Comparison of using multivalent metal salt zinc sulfate solution as protein precipitant and commercially available protein precipitant using existing methods to extract DNA:

[0048] The protein precipitant of the present invention: add zinc sulfate (Sigma-Aldrich China, classification number: Z4750) into deionized water, fully dissolve and prepare solutions with solute solubility of 0.5M, 1M and 2M for later use.

[0049] The commercially available protein precipitant is Buffer AP2 in the AxyPrep Blood Genomic DNA Miniprep Kit (Aisijin Biotechnology (Hangzhou) Co., Ltd., catalog number: AP-MN-BL-GDNA-50).

[0050] Existing methods to extract DNA from whole blood:

[0051] Whole blood samples were stored at 4°C and used within 96 hours after collection. The numbers for each sample are: Y465, Y466, Y467, 2039, 2041, 2045.

[0052] DNA was extracted according to the method of AxyPrep Blood Genomic DNA Miniprep Kit (Aisijin Biotechnology (Hangzhou) Co., Ltd., classificat...

example 2

[0063] The standard method that existing commercially available kit extracts and quick method of the present invention and use polyvalent metal salt zinc sulfate solution as the comparison of protein precipitation agent:

[0064] After the whole blood sample was collected, it was stored at 4°C until it was used, and the DNA was extracted within 96 hours. The numbers of each sample were: Y465, Y466, Y467, 2039, 2041, and 2045. AxyPrep blood genomic DNA miniprep kit and the extracted DNA of the present invention were used respectively. The components of the commercially available kit (Aisijin Biotechnology (Hangzhou) Co., Ltd., catalog number: AP-MN-BL-GDNA-50) include: BufferAP1, BufferAP2, Buffer W1A, BufferW2, Buffer TE and silica gel column. This utility zinc sulfate solution replaces BufferAP2.

[0065] a. The method of extracting DNA with a commercially available kit:

[0066] Add 200ul whole blood sample to a 1.5ml centrifuge tube containing 500ul Buffer AP1, shake vigo...

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PUM

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Abstract

The invention relates to the technical field of biology, in particular to a method for quickly extracting DNA (deoxyribonucleic acid). The method for extracting DNA is chip and easy to operate and comprises the following steps: a, sequentially adding protein precipitate reagent, pyrolytic reagent and a biological sample containing DNA into a container, and uniformly mixing to release cell contents, and agglomerating proteins to obtain lysate which contains the DNA component and non-DNA component released from the biological sample, wherein the protein precipitate reagent is polyvalent metal salt; and b, separating the DNA and non-DNA components in the lysate obtained in the step a to obtain the purified DNA solution. The method has the advantages: protein contamination can be removed by adopting a high-efficiency protein precipitate reagent, the cell lysis and the protein precipitate are synchronously performed. Compared with the prior art, the method has simpler steps and high efficiency, and the use cost is further reduced.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for rapidly extracting DNA. Background technique [0002] In the living body, nucleic acid and protein are the most important substances of life. Nucleic acids carry genetic information and direct the synthesis of proteins. Their relationship is closely linked. When nucleic acid is extracted from some biological samples, such as animal cells, serum, blood, etc., for genetic research or clinical testing at the nucleic acid level, the main problem often encountered is how to remove the abundant protein. [0003] An efficient method known to those skilled in the art is phenol / chloroform extraction, but this method is very time and labor consuming. [0004] An alternative method is to use SDS, protein, and K ions to form a water-insoluble complex, and remove the protein-containing precipitate by centrifugation. This method is commonly used to extract plasmids, the so-called ...

Claims

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Application Information

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IPC IPC(8): C12N15/10
Inventor 龚敬文梁少明任维
Owner 亚能生物技术(深圳)有限公司
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