Method of extracting high-quality DNA (Deoxyribonucleic Acid) of fin product

A high-quality, shark-fin technology, applied in the field of molecular biology-related research, can solve the problems of low and unsatisfactory A260/A230 ratio, and achieve the effect of high DNA purity

Active Publication Date: 2014-08-06
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Through DNA purity analysis, the A260 / A230 ratio of the DNA obtained by this method is low, only about 0.5 (the A260 / A230 ratio of high-purity DNA is usually about 2.0), which cannot meet the requirements of PCR experiments.

Method used

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  • Method of extracting high-quality DNA (Deoxyribonucleic Acid) of fin product
  • Method of extracting high-quality DNA (Deoxyribonucleic Acid) of fin product
  • Method of extracting high-quality DNA (Deoxyribonucleic Acid) of fin product

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Experimental program
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Effect test

Embodiment 1

[0031] (1) Pretreatment of shark fin product samples, including softening shark fins and crushing shark fins in a weak alkaline solution;

[0032]The material used is a sample of shark fin products with the trade name Qingji.

[0033] Use sterilized scissors to cut about 200 mg of shark fin product samples along the shark fin lines from the base to the end, and place them in a 50 mL centrifuge tube. Add approximately 50 mL of sterile 0.06M NaOH solution to the centrifuge tube. Place the centrifuge tube in a shaking water bath and shake at 60°C for 2 hours until the shark's fin is fully softened.

[0034] Centrifuge at 4000rpm for 10 minutes and carefully discard the supernatant. Then, shred the shark's fin and place it in a petri dish, and put it into a vacuum freeze dryer for freeze-drying. The freeze-drying temperature is -54° C., and the freeze-drying time is 8 hours. The freeze-dried samples of shark fin products were ground.

[0035] (2) Lysis and digestion

[0036] ...

Embodiment 2

[0055] (1) Pretreatment

[0056] Pretreatment of shark fin product samples, including softening and crushing shark fins in weak alkaline solution;

[0057] The material used is a sample of shark fin products with the trade name Wuyangfin.

[0058] Use sterilized scissors to cut about 200 mg of shark fin product samples along the shark fin lines from the base to the end, and place them in a 50 mL centrifuge tube. Add approximately 50 mL of sterile 0.06 M NaOH solution to the centrifuge tube.

[0059] Place the centrifuge tube in a shaking water bath, shake the water bath at 60°C for 2 hours, until the shark's fin is fully softened. Centrifuge at 4000rpm for 10 minutes and discard the supernatant carefully.

[0060] After the softened shark's fin is shredded, it is placed in a petri dish, and put into a vacuum freeze dryer to freeze-dry. The freeze-drying temperature is -54°C, the freeze-drying time is 8 hours, and the freeze-dried shark fin product samples are ground.

[0...

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Abstract

The invention relates to a method of extracting high-quality DNA (Deoxyribonucleic Acid) of a fin product and belongs to the field of molecular biology associated research. The method comprises the following steps: (1) pre-treating a fin product sample, namely softening and smashing fins in a weak base solution; (2) splitting the pre-treated fin product sample by a CTAB (Cetyltrimethyl Ammonium Bromide) splitting solution; (3) extracting the DNA by balanced phenol and chloroform-isoamylol in steps; (4) adding a high-saline solution of isopropanol doubled in volume into an aqueous phase containing the DNA to obtain a DNA precipitate; washing the DNA precipitate by an ethanol solution with the volume fraction of 75%, fully drying, and then, dissolving the DNA precipitate and storing. The method of extracting the DNA of the fin product provided by the invention can be used for conveniently and efficiently obtaining the high-quality DNA of a fin tissue so as to avoid pollution by polysaccharide, collagen and other impurities.

Description

technical field [0001] The invention belongs to the field of research related to molecular biology, and in particular relates to a method for extracting DNA from high-quality shark fin products. Background technique [0002] Shark fin is a traditional high-value aquatic product in China, rich in protein, active polysaccharides and mineral elements. Shark fins are processed from shark fins through multiple processing procedures. The sales in the market are often differentiated by the size, origin and processing methods of shark fins. For example, shark fins with a lower degree of processing are usually called green fins. The shark's fin after the cartilage is removed from the green fin is called bright fin, which is the main commercial form of shark fin in the existing aquatic product market. Products such as chrysanthemum fin and wing velvet are all further processed from bright fin. [0003] Recently, the shark fin industry has received a lot of attention around the world....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
Inventor 姚琳王联珠隋哲江艳华李风铃朱文佳郭莹莹翟毓秀
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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