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Mandarin fish paternity test kit and microsatellite PCR identification method thereof

A paternity test and kit technology, applied in the field of molecular biology, can solve the problems of larvae survival impact, larvae cannot be marked, etc., achieve the effect of improving accuracy, improving efficiency, and overcoming interpretation errors

Active Publication Date: 2018-09-28
FRESHWATER FISHERIES RES CENT OF CHINESE ACAD OF FISHERY SCI
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AI Technical Summary

Problems solved by technology

Physical marking can be used for individual identification, but this method has great limitations for juvenile fish, either because the larvae are too small to be marked, or because the marking has a greater impact on the survival of the larvae

Method used

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  • Mandarin fish paternity test kit and microsatellite PCR identification method thereof
  • Mandarin fish paternity test kit and microsatellite PCR identification method thereof
  • Mandarin fish paternity test kit and microsatellite PCR identification method thereof

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Embodiment 1

[0025] Mandarin fish paternity test kit, the kit includes primers for amplifying 20 microsatellite sites, the 20 microsatellite sites are: S26, S27, S29, S31, S32, S34, S38, S44, S50, S57 , S62, S82, S91, S95, S97, S99, S51, S64, S69, S96.

[0026] The primer sequences of the microsatellite loci are: S26, SEQ ID NO.1-2; S27, SEQ ID NO.3-4; S29, SEQ ID NO.5-6; S31, SEQ ID NO.7-8 ; S32, SEQ ID NO.9~10; S34, SEQ ID NO.11~12; S38, SEQ ID NO.13~14; ​​S44, SEQ ID NO.15~16; S50, SEQ ID NO.17~18 ; S57, SEQ ID NO.19-20; S62, SEQ ID NO.21-22; S82, SEQ ID NO.23-24; S91, SEQ ID NO.25-26; S95, SEQ ID NO.27-28; S97, SEQ ID NO.29~30; S99, SEQ ID NO.31~32; S51, SEQ ID NO.33~34; S64, SEQ ID NO.35~36; S69, SEQ ID NO.37~38; S96, SEQ ID NO.39-40. The details of microsatellite loci and primer sequences are shown in Table 1.

[0027] Application of the mandarin fish paternity test kit described above in the parent-child test of mandarin fish, family management or release effect monitoring.

[...

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Abstract

The invention discloses a mandarin fish paternity test kit and a microsatellite PCR identification method thereof. The kit comprises primers which amplify 20 microsatellite loci. The mandarin fish paternity test kit and the microsatellite PCR identification method thereof use the combination of microsatellite markers and a PCR technology to screen the 20 highly polymorphic microsatellite loci, andconduct high-throughput individual recognition and parent-child relationship analysis of the mandarin fish family. A single PCR reaction can detect 4 or 5 sites, relatively simple unit point detection is performed, the efficiency is increased by more than 4 times, and the cost is reduced to about one-third of the original. The error in the interpretation of the intermediate gene size by polyacrylamide gel electrophoresis is overcome by parting through a sequenator, the accuracy of genotype data is improved, and the analysis result shows that 99% of the individuals to be tested can find male and female parents correctly. The establishment of the mandarin fish paternity test kit and the microsatellite PCR identification method thereof provides a new technical means for the mandarin fish germplasm identification, family management and evaluation of the effects of multiplication and emission.

Description

technical field [0001] The invention belongs to the field of molecular biology, and relates to a technology for paternity identification of mandarin fish by using microsatellite marker technology, in particular to a kit for paternity identification of mandarin fish and a microsatellite PCR identification method thereof. Background technique [0002] Mandarin fish (Siniperca chuatsi), also known as Siniperca chuatsi, is a high-quality freshwater cultured fish in China and a unique carnivorous fish. It has the advantages of fast growth, easy feeding, and high nutritional value. With the development and expansion of the mandarin fish farming scale, some farmers have neglected the selection and conservation of the mandarin fish parents, resulting in serious inbreeding of the mandarin fish, which has seriously degraded the germplasm resources of the mandarin fish breeding population and caused disease resistance. Decrease in ability. In order to prevent the degradation of germpl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12Q1/686
CPCC12Q1/686C12Q1/6888C12Q2525/151C12Q2537/143C12Q2563/107
Inventor 丁炜东邴旭文颜元杰曹哲明
Owner FRESHWATER FISHERIES RES CENT OF CHINESE ACAD OF FISHERY SCI
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