Paternity test method and kit of common suckers

A technology of paternity testing and mullet fish, which is applied in the direction of biochemical equipment and methods, microbe determination/testing, etc., to achieve the effect of protecting genetic diversity, fast recovery and multiplication

Inactive Publication Date: 2013-09-04
YANGTZE RIVER FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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Problems solved by technology

However, if the artificial breeding and release work is carried out without considering the genetic background of the species, it will be blind and random, and the artificial breeding and releasing individuals will have a short-term impact on the genetic diversity and population structure of the natural population. impact and long-term effects, it is urgent to use reliable means such as molecular markers to evaluate

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  • Paternity test method and kit of common suckers
  • Paternity test method and kit of common suckers
  • Paternity test method and kit of common suckers

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Embodiment Construction

[0018] Below in conjunction with embodiment, the present invention will be further described.

[0019] 1) DNA extraction of mullet individual

[0020] The tail fin samples of 10 parents and 30 offspring were collected and stored in absolute alcohol at -20°C for later use. Take about 0.5 g of caudal fin tissue, wash the alcohol attached to the caudal fin with double distilled water, soak in TE buffer for 6 hours to fully replace the alcohol, put it into a 1.5mL centrifuge tube and cut it into pieces, add 600μL TE buffer respectively, 40μL 10% SDS, 10μL 20mg / mL proteinase K, mix well, digest at 50℃~54℃ for more than 4 hours, until the tissue is completely digested, add 650ul volume ratio of 25:24:1 phenol-chloroform-isoamyl Alcohol mixture, slowly rotate and mix for 20min, centrifuge at 12000rpm for 10min, absorb the supernatant, repeat once, then add 500ul of chloroform-isoamyl alcohol mixture with a volume ratio of 24:1, mix for more than 20min, centrifuge at 12000rpm for...

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Abstract

The invention discloses a paternity test method of common suckers. The method comprises the following steps of: (1) extracting DNA from common sucker individuals; (2) screening microsatellite markers of the common suckers; (3) carrying out synthesizing and PCR (polymerase chain reaction) amplification on the microsatellite fluorescence markers of the common suckers; and (4) creating paternity test technology of the common suckers. A paternity test kit of the common suckers comprises 10mM / L of 10*PCR Buffer, 25mmol / L of MgCl2, 10mM / L of dNTPs, 10mM / L of 17 microsatellite markers (F.R), 5U / microliter of rTaq Enzyme and ddH2O. The paternity test method and the kit of common suckers disclosed by the invention can be used for realizing artificial reproduction of common sucker farms, providing reference for creating a common sucker family tree, guiding the artificial fecundation and release of the common suckers and improving the reproduction survival rate of the common suckers.

Description

technical field [0001] The invention belongs to the field of aquaculture, and in particular relates to a method and a kit for paternity testing of fish. Background technique [0002] Rouge ( myxocyprinus asiaticus ), Cyprinidae, Substomyidae, and the genus Mummified Fish, this species is the only substomachidae known to be distributed in my country so far. It is distributed in the Yangtze River and Minjiang River systems. Before the 1970s, the mullet was once a One of the large economic fishes in the upper reaches of the Yangtze River. Due to overfishing, water pollution, water conservancy construction, especially after the closure of Gezhou Dam, its resources have declined sharply. At present, it is difficult to find traces of mullet in the Minjiang River. The Yangtze River mullet may be the existing Chinese mullet. The only wild population of mullet is listed as a national second-class wild animal protection. [0003] Microsatellites, also known as short tandem repeats (Sh...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 成为为汪登强危起伟杜浩王成友周琼
Owner YANGTZE RIVER FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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