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Method for extracting abundant total DNA (deoxyribonucleic acid) from grape fruits

A fruit and grape technology, applied in the direction of DNA preparation, recombinant DNA technology, etc., can solve the problems of unfavorable nucleic acid purification, low extraction efficiency, lack of necessary steps, etc., and achieve stable experimental results, good repeatability, and clear agarose electrophoresis bands Effect

Inactive Publication Date: 2015-10-14
江苏绿之邦生态庄园有限公司 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These substances often form complexes with nucleic acids or are oxidized during the extraction process, which is not conducive to the purification of nucleic acids
Therefore, when extracting the total DNA of fruits, the general method does not take into account the above problems, so many necessary steps are missing, resulting in a decrease in extraction efficiency
[0005] The invention provides a method for extracting total DNA from grape fruit, aiming at problems such as polyphenols and polysaccharides in grape fruit and excessive moisture in the ripening period, to efficiently and quickly extract total DNA in grape fruit

Method used

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  • Method for extracting abundant total DNA (deoxyribonucleic acid) from grape fruits
  • Method for extracting abundant total DNA (deoxyribonucleic acid) from grape fruits
  • Method for extracting abundant total DNA (deoxyribonucleic acid) from grape fruits

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] (1) Take 10g of fresh or frozen grape fruit and grind it fully with liquid nitrogen;

[0043] (2) Add to a pre-cooled 50ml centrifuge tube, add 20ml washing buffer (0.1M Tris-boric acid (pH7.4), 0.35M sorbitol, 10% PEG6000, 2% mercaptoethanol) to the centrifuge tube;

[0044] (3) Shake evenly, centrifuge at 8000rpm for 10min at 4°C;

[0045] (4) Remove the supernatant, add about 15ml CTAB buffer (0.1M Tris-boric acid (pH7.4), 2% CTAB, 1.4M NaCl, 20mmol / l EDTA (pH8.0), 2% mercaptoethanol );

[0046] (5) Incubate at 55°C for 30 minutes (CTAB buffer can be preheated in advance), shake by hand several times during incubation;

[0047] (6) Add 1.5ml of 5M KAc, 1.5ml of absolute ethanol, and 15ml of chloroform;

[0048] (7) Place in a shaker at room temperature for 20 minutes, and centrifuge at 1000 rpm for 10 minutes;

[0049] (8) The supernatant was transferred to a new centrifuge tube, and an equal volume of chloroform was added;

[0050] (9) Put it into a shaker at r...

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Abstract

The invention relates to a method for extracting abundant total DNA (deoxyribonucleic acid) from grape fruits, which comprises the following steps: 1. grinding grape fruits into granules; 2. extracting the granules with sorbitol, and filtering out polysaccharides to obtain grape pomace 1; 3. extracting the grape pomace 1 with mercaptoethanol, and filtering out phenols to obtain grape pomace 2; and 4. extracting the grape pomace 2 with a chloroform-isoamyl alcohol-KAc mixture, and filtering out proteins to obtain grape pomace 3 which is the total DNA in the grape fruits.

Description

Technical field: [0001] The invention relates to a method for extracting plant DNA from plants, in particular to a method for extracting a large amount of total DNA from grape fruit. Background technique: [0002] As an important fruit tree species, grapes often require related research. In the process of studying grape molecular biology, there is a considerable need to extract DNA. DNA extraction is the basis for molecular experiments such as gene bank construction, gene cloning, genetic transformation, and molecular markers. Such a common test procedure plays a very important role for further tests in the later stage. The yield, quality, and extraction time of extracted DNA have a great influence on the accuracy and reliability of the experiment. [0003] In the method of extracting the total DNA of plants, people generally use extraction methods common to all plants, such as CTAB method and SDS method, etc. at present. The steps of these extraction methods are basical...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
Inventor 邓洋陈志勇贾海锋房经贵施乃溶朱传根施泽波张荣
Owner 江苏绿之邦生态庄园有限公司
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