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Rapid sex identification primers and kit for trachinotus ovatus and application thereof

A technology of egg-shaped pomfret and a kit, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., can solve the problems of inability to achieve large-scale identification, anatomical individual death, and high equipment requirements. To achieve the effect of shortening the detection cycle, reducing the detection cost and improving the sensitivity

Pending Publication Date: 2021-01-19
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

[0004] (1) In scientific research work, the biological sex of pomfret can be identified through dissection and other methods, but dissection will inevitably lead to the death of the dissected individual, and this method is inefficient and cannot achieve large-scale identification
[0005] (2) The current biotechnology can identify gender in vivo, but the detection technology used is capillary electrophoresis, which requires high equipment, high cost, and cumbersome steps

Method used

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  • Rapid sex identification primers and kit for trachinotus ovatus and application thereof
  • Rapid sex identification primers and kit for trachinotus ovatus and application thereof
  • Rapid sex identification primers and kit for trachinotus ovatus and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0032] The primers for the rapid identification of the sex of the oval pompano provided in this example are primers P18-1, and the primers P18-1 include forward primers P18-1-F and reverse primers P18-1-R, wherein the forward primers The base sequence of P18-1-F is shown in SEQ ID NO: 1, and the base sequence of the reverse primer P18-1-R is shown in SEQ ID NO: 2.

[0033] Specifically, the base sequence of primer P18-1 is as follows:

[0034] Forward primer P18-1-F: 5'-ATGGGACACAGACCCTTCCT-3';

[0035] Reverse primer P18-1-R: 5'-CACCCAGCAGCATAAACAAAT-3'.

[0036] A total of 34 individuals of Pompano ovata were collected, including 16 females and 16 males, and their sexes were identified by anatomy. The sex-determining interval sequence of the amberjack (NCBI accession number AP018739.1) was compared to the reference sequence of the pomfret genome, and an interval was located. Select one of the collected male and female individuals for next-generation sequencing, detect the...

Embodiment 2

[0041] The method for quickly identifying the sex of the oval pomfret provided in this example comprises the following steps: using PCR amplification to amplify specific fragments in the genomic DNA of male and female individuals, specifically including: designing the primers in Example 1, according to the example The method in 1 extracts the genomic DNA of the ovoid pomfret to be tested, adopts the primers in Example 1, and uses the extracted genomic DNA as a template to carry out PCR amplification, adopts agarose gel electrophoresis to detect the amplified product, and according to the electrophoresis result Identify male and female individuals.

[0042] The implementation object of this embodiment is pomfret ovata. Take 50 females and 50 males. The sexes are identified through anatomy, and then the feasibility of the primers is verified.

[0043] (1) Extraction of DNA

[0044] Cut about 1 square centimeter of fin rays from the live pomfret (this process has no substantial ...

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PUM

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Abstract

The invention discloses rapid sex identification primers for trachinotus ovatus, which are primers P18-1, including a forward primer P18-1-F and a reverse primer P18-1-R, the base sequence of the forward primer P18-1-F is shown as SEQ ID NO: 1, and the base sequence of the reverse primer P18-1-R is shown as SEQ ID NO: 2. The invention also discloses a kit including the primers, a method of rapid sex identification for trachinotus ovatus by using the primers, and application of the primers, the kit and the method in rapid sex identification for trachinotus ovatus. The primers have good specificity, good sensitivity and high accuracy, the identification success rate is as high as 97%, and compared with a capillary electrophoresis identification method, the method does not require complicatedequipment, shortens the detection period, and reduces the detection cost.

Description

technical field [0001] The invention belongs to the technical field of ovoid pomfret, and in particular relates to a primer, a kit and an application thereof for rapid gender identification of ovate pomfret. Background technique [0002] Oval pomfret (Trachinotus ovatus), belongs to Perciformes, Trevalidae, Pomfret genus, commonly known as golden pomfret, yellow pomfret, yellow waxed pomfret. Since 2000, large-scale farming has begun in my country, and it has developed into an important marine aquaculture species in China. It is the best species for large-scale aquaculture in deep and open seas. In 2016, the domestic production has reached 120,000 tons. Although its breeding scale has developed rapidly, its breeding work is relatively lagging behind. There are no morphological differences between the male and female individuals of Pomfret ovata in the early embryonic development and juvenile stages, and the male and female cannot be identified from the body size and surface...

Claims

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Application Information

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IPC IPC(8): C12Q1/6879C12Q1/686C12N15/11
CPCC12Q1/6879C12Q1/686C12Q2600/124C12Q2565/125Y02A40/81
Inventor 郭梁张殿昌刘宝锁朱克诚郭华阳杨静文马启伟
Owner SOUTH CHINA SEA FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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