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A method for rapid identification of ivory-participating sea cucumbers

A rough sea cucumber and ivory technology, applied in the field of species identification, can solve the problems of feature disappearance, difficulty in identifying sea cucumber species, and morphological destruction

Active Publication Date: 2020-06-19
MUDANJIANG YOUBO PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the traditional morphological identification method is very susceptible to the influence of many factors such as the environment and genetics during the growth of the species itself; and the morphology is severely damaged or the characteristics disappear during the processing process.
Therefore, it is very difficult to identify sea cucumber species using traditional morphological methods, and it is urgent to establish other effective methods to quickly identify sea cucumber species in the market

Method used

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  • A method for rapid identification of ivory-participating sea cucumbers
  • A method for rapid identification of ivory-participating sea cucumbers
  • A method for rapid identification of ivory-participating sea cucumbers

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Experimental program
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Effect test

Embodiment 1

[0027] 1. Collection of sea cucumber specimens: M1-dried ivory sea cucumber; M2-dried rough sea cucumber; M3-collected sea cucumber samples from random markets

[0028] 2. Pretreatment of test samples: first step, first replace the alcohol in the sea cucumber alcohol-preserved samples and soak the dried sea cucumber samples. The solutions used are PBS (137mmol / L NaCl, 2.7mmol / L KCl, 10mmol / L Na2HPO4 , 2mmol / LKH2PO4, pH 7.2) In the second step, take 50-80mg of the sample processed in the previous step and put it in 800-1000μl high-salt buffer (200mmol / L Tris-HCl, 50mmol / L EDTA, 500mmol / L NaCl, 0.2 % β-mercaptoethanol, pH 8.0), centrifuge at 10000-12000g / min for 1-2min, discard the supernatant; repeat the operation twice. Proteinase K was incubated at 65°C for 1 hour until the tissue was completely dissolved. Add an equal volume of chloroform:isoamyl alcohol (24:1) and mix well, centrifuge at 15,000×g for 10 minutes at room temperature, and carefully pipette the supernatant int...

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Abstract

The invention discloses a DNA (Deoxyribonucleic Acid) barcode molecular identification method of two types of sea cucumbers (holothuria fuscopunctata and holothuria scabra) and a CO I sequence of the sea cucumbers. The method is used for carrying out PCR (Polymerase Chain Reaction) amplification to obtain a CO I gene of the sea cucumbers; a PCR product is identified through agarose gel and then is sequenced; a sequencing result is compared with a public sequence; if the homology between the sequencing result and a gene sequence SEQ ID NO.1 is 99 percent or above, the method can judge that a tissue to be detected is a source of the holothuria fuscopunctata; if the homology between the sequencing result and a gene sequence SEQ ID NO.2 is 99 percent or above, the method can judge that the tissue to be detected is the holothuria scabra. According to the DNA barcode molecular identification method, a reliable DNA technology is adopted, and holothuria fuscopunctata and holothuria scabra varieties are rapidly and accurately distinguished; the accuracy and reliability are enhanced, and the safety of taking the sea cucumbers as a medicinal material is ensured.

Description

technical field [0001] The invention relates to the field of species identification, in particular to a DNA barcode molecular identification method for two kinds of sea cucumbers (ivory sea cucumber and rough sea cucumber). Background technique [0002] Sea cucumber is the general term for animals of the class Holothrioider in the phylum Echinodermata. It is a traditional sea-like food and drug resource. . The prices of different species of sea cucumbers vary greatly, but many economic species of sea cucumbers lack clear species identification during the trade process. Ivory ginseng (Holothuria fuscopunctata) is usually 6-30cm long and 0.8-10cm wide. The whole surface is grayish white or yellowish white, wrinkled or horizontally striated, and some horizontally wrinkled deeply into grooves, and the grooves are yellow or brown. . The color is darker in the middle of the back, lighter on both sides, and gradually turns white on the ventral surface; there is often an obvious ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12Q1/6869C12N15/53
CPCC12Q1/6869C12Q1/6888C12Q2531/113C12Q2563/185
Inventor 李振国陈艳明务勇圣郭晓璐
Owner MUDANJIANG YOUBO PHARMA CO LTD
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