Method for rapidly identifying genders of macrobrachium rosenbergii

A technology of Macrobrachium rosenbergii and sex, applied in the field of aquaculture, can solve the problems of inability to distinguish ZW and WW genotypes, difficult to accurately judge gender, etc., and achieve the effect of high detection sensitivity

Active Publication Date: 2020-05-08
深圳市泽龙生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the lack of male-specific genomic fragments, this method has certain limitations: 1. ZW and WW genotypes cannot be distinguished; 2. False positives (such as PCR contamination) or false negatives (genome quality problems or problems with PCR amplification) ) is difficult to accurately determine gender

Method used

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  • Method for rapidly identifying genders of macrobrachium rosenbergii
  • Method for rapidly identifying genders of macrobrachium rosenbergii
  • Method for rapidly identifying genders of macrobrachium rosenbergii

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Embodiment 1

[0057] Amplified length polymorphism amplification of the Macrobrachium rosenbergii genome was performed at a lower annealing temperature (Jiang et al., 2013), and the sex-specific genome fragments of the Macrobrachium rosenbergii were obtained.

[0058] 1. Extraction of Macrobrachium rosenbergii Genomic DNA

[0059] Cut the tentacles of Macrobrachium rosenbergii, and extract the genomic DNA by the phenol-chloroform method:

[0060] Take 4 giant giant macrobrachium rosenbergii female shrimp (ZW). Disinfect the tweezers, cut about 2-3 tentacles, and digest with tissue lysate and proteinase K (600 μL in total) until the tissue is completely dissolved. Add an equal volume of phenol: chloroform: isoamyl alcohol (25:24:1) solution, mix well and centrifuge. Transfer the supernatant to a new centrifuge tube and add an equal volume of pre-cooled absolute ethanol, mix well and centrifuge, discard the supernatant. Wash the precipitate with 70% ethanol, discard the ethanol and air dry...

Embodiment 2

[0067] This embodiment provides a method for quickly identifying the sex of Macrobrachium rosenbergii, comprising the following steps:

[0068] 1. Extraction of Macrobrachium rosenbergii Genomic DNA

[0069] Cut the tentacles of Macrobrachium rosenbergii, and extract the genomic DNA by the phenol-chloroform method:

[0070] Four male Macrobrachium rosenbergii males (ZZ), four females (ZW) and super females (WW) were taken respectively. Disinfect the tweezers, cut about 2-3 tentacles, and digest with tissue lysate and proteinase K (600 μL in total) until the tissue is completely dissolved. Add an equal volume of phenol: chloroform: isoamyl alcohol (25:24:1) solution, mix well and centrifuge. Transfer the supernatant to a new centrifuge tube and add an equal volume of pre-cooled absolute ethanol, mix well and centrifuge, discard the supernatant. Wash the precipitate with 70% ethanol, discard the ethanol and air dry the precipitate; add 20-50 μL TE buffer to detect the DNA con...

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Abstract

The invention is applicable to the technical field of aquaculture and in particular relates to a method for rapidly identifying genders of macrobrachium rosenbergii. The method comprises the followingsteps: extracting macrobrachium rosenbergii genomic DNA (deoxyribonucleic acid) from macrobrachium rosenbergii tissue; designing primers, and performing PCR (polymerase chain reaction) amplification,wherein the primer pairs have sequences shown in SEQ ID NO:3 and SEQ ID NO:4 or sequences with 80% or greater of homology with SEQ ID NO:3 and SEQ ID NO:4; and performing electrophoretic separation on a product obtained after PCR amplification, and judging genetic genders of detected macrobrachium rosenbergii according to banding patterns. By adopting the method, macrobrachium rosenbergii of three different genotypes (ZZ, ZW and WW) can be rapidly distinguished through one time of PCR by using one primer pair, genders of parent shrimps and shrimp seeds of unisexual fries can be identified, and great application value can be achieved.

Description

technical field [0001] The invention belongs to the technical field of aquaculture, and in particular relates to a method for quickly identifying the sex of Macrobrachium rosenbergii. Background technique [0002] Macrobrachium rosenbergii (Macrobrachium rosenbergii) belongs to the phylum Arthropoda, grows in freshwater or brackish water, and is distributed in tropical and subtropical regions of the Indian Ocean and Pacific Ocean. It has a large body, a wide range of food habits, fresh and tender meat, and high nutritional value. It is one of the shrimp species with high breeding value in the world. [0003] The breeding process of Macrobrachium rosenbergii has the following characteristics: 1. There is a big difference in the growth of male and female shrimp, and the growth rate of male shrimp is more than 50% faster than that of female shrimp; 2. The male and female shrimp of Macrobrachium rosenbergii are mixed, and sexual maturity starts to mate and consumes energy to aff...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6879
CPCC12Q1/6879C12Q2600/156Y02A40/81
Inventor 刘云杨玲玲
Owner 深圳市泽龙生物技术有限公司
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