Method for direct extraction of nucleic acid without tissue grinding

An extraction method and nucleic acid extraction technology, which are applied in the fields of biochemical equipment and methods, recombinant DNA technology, and the determination/inspection of microorganisms to achieve the effects of fast extraction, stable results and easy operation.

Inactive Publication Date: 2017-09-01
BIOTEKE CORP (WUXI) CO LTD
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0013] The present invention needs to overcome the defect in the traditional nucleic acid extraction process:

Method used

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  • Method for direct extraction of nucleic acid without tissue grinding
  • Method for direct extraction of nucleic acid without tissue grinding
  • Method for direct extraction of nucleic acid without tissue grinding

Examples

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Embodiment 1

[0039] 1. Mouse liver DNA extraction

[0040] Manual reagent addition steps (if it is a sealed kit, add 125 μl lysate TR to each well of the first 96 deep-well plate, add 20-60 mg of mouse liver, 20 μl proteinase K, 4 μl RNase, and then directly on the computer):

[0041] 1) Add 250 μl magnetic bead binding solution CB, 125 μl lysis solution TL and 125 μl lysis solution TR, 20 μl proteinase K, 4 μl RNase and about 10 mg tissue sample to the first 96 deep-well plate;

[0042] 2) Add 300 μl buffer BB and 20 μl magnetic beads to each well of the second deep well plate;

[0043] 3) Add 900 μl inhibitor remover IR to each well of the third and fourth deep well plates;

[0044] 4) Add 900 μl of washing solution WB to each well of the fifth deep well plate;

[0045] 5) Add 150 μl of elution buffer TE to each well of the sixth deep well plate;

[0046] 6) Place the deep-well plates in the positioning seats of the 96 nucleic acid extractor in sequence, and then insert the stirring ...

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Abstract

The invention relates to the field of nucleic acid extraction, in particular to a method for direct extraction of nucleic acid without tissue grinding. In order to solve the defects of burdensome operation, long extraction cycle, no realization of automation, contact of toxic reagents and the like, and the technical problems that in the extraction process of a novel nucleic acid extraction instrument, animal tissue samples need grinding, cracking and centrifugation in advance and cannot achieve direct extraction on a machine, the invention provides a method for direct extraction of nucleic acid without tissue grinding. The method adopts a unique cracking reagent and employs a 96-channel nucleic acid extraction instrument for support to realize integration of instrument and consumables, and achieves full-automatic extraction of animal tissue nucleic acid. The reagent is optimized cracking liquid for different samples, and the instrument is nucleic acid extraction instrument, and the extraction conditions can be optimized according to different tissue samples. The method provided by the invention has the advantages of simple steps, full automatic operation, high purity of nucleic acid, short extraction cycle, and no contact with toxic reagent, etc.

Description

technical field [0001] The invention belongs to the technical field of nucleic acid extraction, and relates to a nucleic acid extraction method, in particular to a method for directly extracting nucleic acid from tissues without grinding. Background technique [0002] Nucleic acid research is an important topic in modern biology and medical research. As the carrier of genetic information, nucleic acid is the material basis of gene expression. In addition to playing a very important role in the normal growth, development, and reproduction of organisms, it is also related to abnormal conditions of life, such as tumor occurrence, radiation, etc. Injuries, genetic diseases, etc. are also closely related. With the advancement of molecular biology techniques in recent years, molecular diagnosis and detection techniques represented by nucleic acid hybridization, nucleic acid amplification, and nucleic acid sequence analysis are increasingly playing a vital role in many fields. Ne...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
CPCC12N15/1013C12Q2523/308C12Q2563/143C12Q2563/149
Inventor 周志图
Owner BIOTEKE CORP (WUXI) CO LTD
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