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Method for simultaneously identifying genetic sexes of litopenaeus vannamei at DNA or RNA level

A technology for vannamei and prawns, applied in the fields of molecular biology and genetic breeding, can solve the problems of undetectable markers, lack of limited development, etc., and achieve the effects of accurate and reliable results, improved adaptability, and simple operation.

Pending Publication Date: 2020-03-17
INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Abstract
  • Description
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  • Application Information

AI Technical Summary

Problems solved by technology

However, this marker is located in a non-coding region of the genome where transcription does not occur, so it cannot be detected at the RNA level
However, when studying sex determination and differentiation, it is very important to sex the extracted individual RNA, and the lack of this technology limits the development of sex research in Litopenaeus vannamei

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0015] Example 1: Identifying the genetic sex of Litopenaeus vannamei by extracting tissue DNA

[0016] (1) Source of material

[0017] The materials of 6 different germplasm resources with known gender collected in the laboratory were used. Each germplasm material had 16 individuals, a total of 96 individuals, and the ratio of male to female was 1:1. Individual DNA was extracted using the Tiangen Plant Genome Extraction Kit, the DNA concentration was determined using a Nanodrop 1000 spectrophotometer, and the integrity of the band was detected by agarose gel electrophoresis.

[0018] (2) PCR amplification and sequencing analysis

[0019] For the above 96 individuals, use Tiangen Golden easy PCR mix (Tiangen, Beijing, China) to amplify. The amplification system is as follows: 2 μl DNA template, 0.5 μl LvDRSMF and 0.5 μl LvDRSMR, 12.5 μl PCR mix, 9.5 μl sterilized Water, the total system is 25ul. The PCR reaction program was as follows: pre-denaturation at 94°C for 3min, (94...

Embodiment 2

[0026] Example 2: Using tissue RNA to identify the genetic sex of Litopenaeus vannamei

[0027] (1) Source of material

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PUM

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Abstract

The invention relates to a method for simultaneously identifying the genetic sexes of litopenaeus vannamei at the DNA or RNA level. The early-stage genetic sex determination of prawns not only is of great significance in a production process, but also plays an important role in researching the molecular mechanisms of sex determination and sex differentiation. Previously reported prawn genetic identification methods can only use the DNAs of individuals to perform molecular identification, and no related method for identifying sexes by using DNAs and RNAs exists at present. According to the method for simultaneously identifying the genetic sexes of litopenaeus vannamei at the DNA or RNA level, three SNP markers with sex differences on sex-related genes are utilized, and the SNPs are typed, and therefore, the sexes of the individuals can be identified through the DNAs or RNAs of the individuals. With the method adopted, the obstacle that sex determination cannot be realized by using the RNAs of individuals in the past can be eliminated; a new sex determination method and means are provided; and a favorable tool is provided for subsequent sex determination and sex differentiation research.

Description

technical field [0001] The invention belongs to the fields of molecular biology and genetic breeding, and in particular relates to a method capable of simultaneously identifying the sex of Litopenaeus vannamei at the levels of DNA and RNA. Background technique [0002] Sex determination is an important biological process in life sciences, and the study of biological sex determination mechanism plays an important role in revealing the evolution of sex. At the same time, because there are obvious differences in growth traits between male and female individuals of many economic animals, the study of sex determination is also of great significance for parthenogenesis control and breeding of parthenogenetic breeds. [0003] Litopenaeus vannamei is the main mariculture species in my country, with an annual output of 1.5 million tons. The male and female individuals of this variety show obvious growth differences in the middle and late stages, and the growth rate of female individ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6879C12Q1/6888
CPCC12Q1/6879C12Q1/6888C12Q2600/156Y02A40/81
Inventor 于洋李富花王悦相建海
Owner INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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