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DNA barcoding standard detection sequence for acrossocheilus beijiangensis and applications of DNA barcoding standard detection sequence

A standard detection, light lip fish technology, applied in the application, microbial determination/inspection, biochemical equipment and methods, etc., can solve the problem of lack of data and other problems, achieve high repeatability, shorten identification time, and good stability.

Inactive Publication Date: 2016-10-05
SHENZHEN HUADA GENE INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the endemism of many fishes in our country, there is not enough relevant data

Method used

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  • DNA barcoding standard detection sequence for acrossocheilus beijiangensis and applications of DNA barcoding standard detection sequence
  • DNA barcoding standard detection sequence for acrossocheilus beijiangensis and applications of DNA barcoding standard detection sequence
  • DNA barcoding standard detection sequence for acrossocheilus beijiangensis and applications of DNA barcoding standard detection sequence

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] 1. Collection, storage and processing of samples to be tested

[0039] Cut part of the muscle tissue of the North River lip fish from the soaked specimen, store it in 95% alcohol and store it in a refrigerator at -20°C.

[0040] 2. DNA extraction

[0041] The DNA in the samples was extracted by the phenol-chloroform method and stored at -20°C for future use.

[0042] 3. Synthesis of PCR primers

[0043] This method selects 2 pairs of general primers for fish:

[0044] FishF2_t1:5'-TGTAAAACGACGGCCAGTCGACTAATCATAAAGATATCGGCAC-3' (SEQ ID NO:2);

[0045] FishR2_t1:5'-CAGGAAACAGCTATGACACTTCAGGGTGACCGAAGAATCAGAA-3' (SEQ ID NO:3);

[0046] VF2_t1: 5'-TGTAAAACGACGGCCAGTCAACCAACCACAAAGACATTGGCAC-3' (SEQ ID NO: 4);

[0047] FR1d_t1:5'-CAGGAAACAGCTATGACACCTCAGGGTGTCCGAARAAYCARAA-3' (SEQ ID NO:5).

[0048] 4. PCR reaction system (25μl)

[0049]

[0050]

[0051] 5. PCR reaction procedure

[0052] 95℃*2min,

[0053]

[0054] 72°C*10min.

[0055] After the reaction...

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Abstract

The invention discloses a DNA barcoding standard detection sequence for acrossocheilus beijiangensis, which is shown as SEQ ID NO;1, and a molecular identification method based on the standard detection sequence. Due to the application of the DNA barcoding standard detection sequence for acrossocheilus beijiangensis provided by the invention, the reliable DNA molecular identification technology for the incomplete individuals which can not be accurately identified and for the complete individuals in samples is realized, therefore, the technical defect that in the prior art, the incomplete individuals basically can not be identified, and the complete individuals need to be identified by the professionals who are very familiar to the morphological traits of acrossocheilus beijiangensis is overcome, and compared with the traditional morphological identification method, the DNA barcoding standard detection sequence provided by the invention can be used for rapidly and accurately identifying acrossocheilus beijiangensis.

Description

technical field [0001] The invention relates to the technical field of species identification, in particular to a DNA barcode standard detection sequence of Beijiang light-lipped fish and its application in species identification. Background technique [0002] Beijiang light-lipped fish (scientific name: Acrossocheilus beijiangensis) is a fish of the genus Acrossocheilus beijiangensis of the order Cyprinidae of the ray-finned fishes, and it is a unique species in China. Distributed in Emei, Jiading, Suji and other places in Sichuan Province, it is common in the main stream of rivers. The type locality of this species is Mount Emei. Distributed in Xijiang and Beijiang of the Pearl River system, Hong Kong also has a very small amount of distribution and is recognized as a rare species. The type locality of this species is Lianzhou and Yangshan in the middle and upper reaches of Lianhe in the Pearl River Basin. Beijiang smooth lip fish lays sticky eggs and spawns on gravel, ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/53
Inventor 王敏孙颖赵俊赵晓萌何舜平刘荭李启沅潘文欢石琼
Owner SHENZHEN HUADA GENE INST
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