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A method for identifying mesogroups and microgroups of kite squid using a specific amplification primer set

A technology of kite squid and primer set, applied in the field of molecular marker identification, can solve problems such as indistinguishability, and achieve the effect of simple operation

Active Publication Date: 2021-09-03
SOUTH CHINA SEA FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the mesogroups and microgroups in the larval stage are similar in appearance, and it is difficult to distinguish them by their appearance

Method used

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  • A method for identifying mesogroups and microgroups of kite squid using a specific amplification primer set
  • A method for identifying mesogroups and microgroups of kite squid using a specific amplification primer set

Examples

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Embodiment 1

[0018] A method for identifying mesogroups and microgroups of kite squid using a specific primer set, the specific primer set is composed of a forward primer MS700F and two reverse primers M800R and MS400R, and the corresponding base sequences are respectively:

[0019] MS700F: 5'-TGACTATTTTCTACAAACCAT-3';

[0020] M800R: 5'-CTACAGTGAACATGTGATGAG-3';

[0021] MS400R: 5'-AACTGATGGTCCAGCAT-3'.

Embodiment 2

[0023] A method for identifying mesogroups and microgroups of kite squids using specific primer sets, comprising the following steps:

[0024] (1) Prepare sample DNA using mesogroup or microgroup of kite squid; preferably, the sample DNA is extracted from 20-50 mg of back muscle of mesogroup or microgroup of kite squid using Marine Animal Genomic DNA Extraction Kit (TIANGEN) The extracted genomic DNA was prepared by dissolving in 100ml double distilled water.

[0025] (2) Perform PCR amplification on the PCR amplification system containing the sample DNA and a specific primer set; preferably, the PCR amplification system: the total volume of the amplification reaction is 30 μl, including: 3 μL 10×buffer (Mg 2+ plus), dNTP Mixture 2.5mM each, primers 0.5μM each, 0.5 U Taq enzyme (TaKaRa) and 3μL template DNA; preferably, the reaction conditions of the PCR amplification are: pre-denaturation at 95°C for 1 min, denaturation at 94°C for 40s , annealing at 48°C for 40 s, extension...

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Abstract

The invention discloses a method for identifying mesogroups and microgroups of kite squid by using a specific primer set. The specific primer set is composed of a forward primer MS700F and two reverse primers M800R and MS400R. The forward primer MS700F is composed of The base sequence is shown in SEQ NO.1, the base sequences of the reverse primer M800R and the reverse primer MS400R are shown in SEQ NO.2 and SEQ NO.3 respectively, and a specific primer set is used to amplify the kite squid by PCR The genomic DNA of the medium group and the microgroup was detected by electrophoresis, and the electrophoresis spectrum showed that only a specific bright band appeared in the PCR amplification product of the kite squid microgroup at nearly 400bp, and the PCR amplification product of the kite squid mesogroup was near 400bp and Nearly 800bp each appeared a specific bright band, so as to accurately and quickly distinguish the two kinds of kite squid.

Description

technical field [0001] The invention relates to the field of molecular marker identification, in particular to a method for identifying mesogroups and microgroups of kite squid by using a specific amplification primer set. Background technique [0002] Kite squid (Sthenoteuthisoualaniensis), commonly known as red squid, belongs to Teuthoidea, Ommastrephidae, and Sthenoteuthis. Purple-brown, with oval luminescent tissue under the skin in front of the back of the carcass, and the 4th wrist on the left side of the male is stalked. The kite squid is a warm-water oceanic species widely distributed in the tropical and subtropical waters of the Indian Ocean and the Pacific Ocean. Among them, the number of kite squid is the most abundant in the South China Sea and the northwestern part of the Indian Ocean, and it is the species with the most potential for development in the oceanic fishery resources of the South my country Sea. . South China Sea kite squid can be divided into two p...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888
CPCC12Q1/6888
Inventor 徐磊杜飞雁王雪辉黎红宁加佳张鹏王亮根李亚芳徐亮
Owner SOUTH CHINA SEA FISHERIES RES INST CHINESE ACAD OF FISHERY SCI