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Microsatellite marker primer pair for identifying genetic sex of brachymystax lenok salmon and sex identification method

A technology of microsatellite marker and identification method, applied in the field of microsatellite marker primer pair and gender identification, can solve the problems of inability to take gonads in all fish, cumbersome process, inability to fully understand the sex ratio of wild fish, etc. Effect

Pending Publication Date: 2022-05-13
HEILONGJIANG RIVER FISHERY RES INST CHINESE ACADEMY OF FISHERIES SCI +1
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  • Summary
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AI Technical Summary

Problems solved by technology

Lepidopteris is a rare and precious cold-water fish with scarce wild resources. In the past, sex identification required taking gonads and making paraffin sections. The process is cumbersome and requires killing fish, and it is impossible to identify small fish with immature gonads.
In view of the fact that the previous method of gender identification required killing fish, but in the process of wild surveys, all fish cannot be taken from gonads to identify sex, especially the lepidoptera is a national second-class protected animal, so it is impossible to fully understand the sex ratio of wild fish

Method used

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  • Microsatellite marker primer pair for identifying genetic sex of brachymystax lenok salmon and sex identification method

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specific Embodiment approach 1

[0015] Specific embodiment 1: This embodiment is used to identify the microsatellite marker primer pair for the genetic sex of leprosy salmon, wherein the sequence of the upstream primer F in the primer pair is 5'-CACCCACACACGTACCAGTC-3', and the sequence of the downstream primer R in the primer pair The sequence is 5'-TGTGGAGTTCATGTGGGATG-3'.

specific Embodiment approach 2

[0016] Specific embodiment two: the method for identification of the genetic sex of leprosy salmon in this embodiment:

[0017] 1. Extracting the DNA of the salmon to be identified;

[0018] 2. PCR reaction; the amplification primers in the PCR reaction system are the microsatellite marker primer pair described in Embodiment 1; the sequence of the upstream primer F is 5'-CACCCACACACGTACCAGTC-3', and the sequence of the downstream primer R is 5'-TGTGGAGTTCATGTGGGATG- 3';

[0019] 3. Agarose gel detection, the amplified result of 0 band is female fish, and the amplified result of 1 band is male fish; the amplified band of male fish is 278bp.

specific Embodiment approach 3

[0020] Specific embodiment 3: The difference between this embodiment and specific embodiment 2 is that in step 2, the PCR reaction system adds a reference primer for the salmon 18s ribosomal RNA as a reference;

[0021] The upstream primer 18SF in the reference primer pair for 18s ribosomal RNA of leprosy salmon is 5'-GGTCCGAAGCGTTTACTTTG-3', and the downstream primer 18SR is 5'-ACCTCTAGCGGCACAATACG-3';

[0022] Step 3, agarose gel detection, if the amplification result is 1 band, it is a female fish, and if the amplification result is 2 bands, it is a male fish. Others are the same as in the second embodiment.

[0023] In this embodiment, the 18s ribosomal RNA gene of the leptospirosis is used as a reference, and whether the PCR amplification result of the reference gene is present or not can determine whether the electrophoretic band loss is caused by reasons such as sample addition errors, amplification failures, and DNA degradation. Improve the accuracy of identification ...

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Abstract

The invention discloses a microsatellite marker primer pair for identifying genetic sex of brachymystax lenok and a sex identification method, and relates to a microsatellite marker primer pair for identifying fish sex and a sex identification method. The sequence of an upstream primer F in a primer pair of the microsatellite marker disclosed by the invention is 5 '-CACCACACCGTACCAGTC-3', and the sequence of a downstream primer R in the primer pair is 5 '-TGTGGAGTGGGATG-3'. The sex identification method comprises the following steps: 1, extracting DNA of brachymystax lenok salmon to be identified; 2, carrying out PCR (Polymerase Chain Reaction) reaction; and 3, agarose gel electrophoresis detection. According to the method, the sex is judged through the agarose gel electrophoresis band of the PCR amplification product, only DNA samples need to be extracted, fin rays or scales can be collected, DNA can be extracted, damage to fish bodies is small, fish killing is not needed, and the method can be used for early sex identification.

Description

technical field [0001] The invention relates to a pair of microsatellite marker primers for fish sex identification and a sex identification method. Background technique [0002] Brachymystax lenok (Pallas), belonging to the order Salmoniformes and the family Salmonidae, Brachymystax is a rare and valuable cold-water fish in China. It was listed in the Red Data Book of Endangered Animals in China in 1998 , the endangered level is endangered, and the wild population will be listed as a national second-class protected animal in 2021. [0003] Lepidoptera is sexually mature at the age of 3 to 5, and has no secondary sexual characteristics when it is sexually mature. It is difficult to identify sex by appearance during the breeding season. Lepidopteris is a rare and valuable cold-water fish with scarce wild resources. In the past, sex identification required taking gonads and making paraffin sections. The process was cumbersome and required killing fish, and it was impossible t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12Q1/6879C12Q1/686C12N15/11
CPCC12Q1/6888C12Q1/6879C12Q1/686C12Q2565/125C12Q2525/151Y02A40/81
Inventor 佟广香刘冰宋建臣刘福祥韩世成马凯匡友谊
Owner HEILONGJIANG RIVER FISHERY RES INST CHINESE ACADEMY OF FISHERIES SCI
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