Pelodiscus sinensis sex identification primer set, locus and method

A technology of Chinese soft-shelled turtle and primer set, which is applied in biochemical equipment and methods, recombinant DNA technology, microbial measurement/inspection, etc., and can solve problems such as long time-consuming and difficult gender

Active Publication Date: 2018-09-28
ZHEJIANG UNIV
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Problems solved by technology

At present, there is no effective molecular method to identify the sex of soft-shelled soft-shelled turtles. However, it is difficult and time-consuming to identify the sex of soft-shelled

Method used

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  • Pelodiscus sinensis sex identification primer set, locus and method
  • Pelodiscus sinensis sex identification primer set, locus and method
  • Pelodiscus sinensis sex identification primer set, locus and method

Examples

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Embodiment Construction

[0021] The Chinese soft-shelled turtle gender identification primer set design and typing of the present invention have three steps: 1. Designing female-specific primers for the difference segment of the sex chromosome linkage gene found in previous experiments; 2. Designing both male and female soft-shelled turtle genomic DNAs. Can effectively amplify, and will not produce positive reference primers for non-specific amplification with female-specific primer pairs; 3. Amplify the adult Chinese soft-shelled turtle genome DNA with the synthesized primer set, check its amplification accuracy and efficiency, and Adjust the ratio of the primer set, annealing temperature, etc.; 4. Use the amplified product in step 3, take pictures with a gel imager, and read the electrophoresis bands to confirm the gender.

[0022] Below in conjunction with example the invention is further described:

[0023] Sample preparation: Individual liver tissues of soft-shelled turtle samples were collected ...

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Abstract

The invention discloses pelodiscus sinensis sex identification specific primers and a method. The two pairs of primers in a sex identification primer system are a pair of female specific primers and apair of positive reference primers which can be amplified by using male and female genomic DNA. Among the primers, the female specific primers can only stably amplify a 196 bp band in the female genomic DNA, and the band cannot be amplified in the male genomic DNA; the positive reference primers can amplify a 482 bp band in the male and female genomic DNA. Thus, in a PCR reaction system, the female genomic DNA can amplify two bands of 196 bp and 482 bp, while the male can only amplify a band of 482 bp. The use of the primers facilitates the correct identification of the sex of pelodiscus sinensis, especially the sex identification of the pelodiscus sinensis in the embryonic stage which is difficult to distinguish through naked eyes. The primers lay the foundation for the study of the pelodiscus sinensis, and also provide an important tool for the breeding and artificial breeding of the pelodiscus sinensis.

Description

technical field [0001] The invention relates to a specific primer for amplifying the specific sequence of the Chinese soft-shelled turtle W chromosome-linked gene SBNO1 and a positive reference primer capable of amplifying a 482bp sequence from both male and female Chinese soft-shelled turtle genome DNA, in particular to a primer set and identification for sex identification of Chinese soft-shelled turtle method. Background technique [0002] Chinese soft-shelled soft-shelled turtle (Pelodiscus sinensis) belongs to the genus Trionychidae of the order Testudinata of the class Reptilia, and is also known as soft-shelled turtle, turtle, water fish, etc. It is an important aquatic reptile. It is listed as a Vulnerable (VU) species on the IUCN Red List of Threatened Species. It is widely distributed in other provinces in my country except Xinjiang, Tibet and Qinghai, and has also been introduced in Vietnam, Japan, Korea, Timor Island and the Hawaiian Islands. [0003] Chinese s...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/6879C12N15/11
CPCC12Q1/6879C12Q1/6888
Inventor 方盛国万秋红王歆
Owner ZHEJIANG UNIV
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