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Molecular marker primer and method for identifying brown spotted flat head fishes and Indian flat head fishes

A molecular marker, molecular marker sequence technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc. Increased or decreased amplification efficiency, high consistency, and stable results

Inactive Publication Date: 2019-01-04
ZHEJIANG OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

There is no formal report on the molecular markers used to identify S.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] A kind of molecular marker primer for distinguishing brown scallops and Indian scallops, the primer sequence is:

[0024] 12SF: 5'-GTCGGTAAAACTCGTGCCAGC-3';

[0025] 12SR: 5'-CATAGTGGGGTATCTAATCCCAGTTTG-3'.

[0026] The length of the molecular marker primer and the difference between the length of the upstream and downstream primers are more suitable for the amplification of the PCR reaction, and the T of the upstream and downstream primers m The difference between the values ​​is within 5°C, which can effectively avoid non-specific amplification or decrease in amplification efficiency, increase the success of the PCR reaction, and improve the accuracy and stability of the identification results.

[0027] A kind of method that utilizes above-mentioned primer to discriminate brown scallop and Indian scallion, concrete steps are as follows:

[0028] 1) Extraction of the DNA of the brown crocodile and Indian crocodile: Take the muscle sample and place it in a 1.5ml centr...

Embodiment 2

[0040] The DNA of 15 samples was extracted from 10 brown scallops collected from Qingdao, Zhoushan and Xiamen populations and 5 Indian scallops collected from Guangzhou population, and then the extracted DNA was used as a template for PCR amplification, and the amplification results were analyzed. Perform purification, sequencing, and compare the sequencing results;

[0041] Among them, the primer sequence is:

[0042] 12SF: 5'-GTCGGTAAAACTCGTGCCAGC-3';

[0043] 12SR: 5'-CATAGTGGGGTATCTAATCCCAGTTTG-3'.

[0044] The composition of the reaction system for PCR amplification is as follows: Add in sequence to a 0.2ml centrifuge tube:

[0045] Ultrapure water 17.5ul

[0046] 10×buffer 2.5ul

[0047] dNTPs 2ul

[0048] rTaq 0.15ul

[0049] Template 1ul

[0050] Primer 1ul each;

[0051] The PCR amplification program was: pre-denaturation at 94°C for 5 min, (denaturation at 94°C for 45 sec, annealing at 50°C for 45 sec, extension at 72°C for 45 sec) × 35 cycles, extension at 7...

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Abstract

The invention discloses a molecular marker primer and method for identifying brown spotted flat head fishes and Indian flat head fishes. The nucleotide sequence of the primer is shown as SEQ ID No.1 and SEQ ID No.2. The identification method specifically includes the steps that DNAs of the brown spotted flat head fishes and DNAs of Indian flat head fishes are extracted, the DNAs of the two kinds of fishes are used as templates for PCR amplification, amplification results are purified and sequenced, and sequencing results are compared. The molecular marker primer and method have the advantagesthat the molecular marker primer is suitable for amplification of a PCR reaction and can effectively avoid non-specific amplification or decline of the amplification efficiency, increase the success rate of the PCR reaction and improve the accuracy and stability of identification results; the identification method is stable in result, high in repeatability and simple, the amplification results ofdifferent individuals are the same, and high consistency is achieved.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a molecular marker primer and a method for distinguishing brown scallops and Indian scallops. technical background [0002] The brown-spotted scorpion, like the Indian scorpion, belongs to the class Teleost, Scorpionidae, Scorpionidae, and Scorpion in taxonomy. Morphologically, the brown-spotted scorpion is very similar to the Indian scorpion, with dorsal fin spines I-I-VI~VII-I, dorsal fin rays 13~14, pectoral fin rays 17~19, pelvic fin spine I, pelvic fin rays 5, anal fin rays 13 ~14, lateral line scales 83~99, lateral line scales 18~21, lateral line inferior scales 30~35. Body elongated, head flattened, tapering posteriorly. The back is dark brown, the abdomen is white, the back is scattered with brown spots, the caudal fin is white, and there are 2~3 black stripes on it. It is mainly distributed in the continental shelf waters of the western Pacific Ocean, and it...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/156
Inventor 高天翔徐胜勇陈治王志杨杨天燕蔡珊珊
Owner ZHEJIANG OCEAN UNIV
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