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A primer, kit and identification method for identifying the genetic relationship among individuals of the Yangtze finless porpoise

A kind of kinship, kit technology, applied in the field of molecular biology, can solve problems such as determining the kinship of groups

Active Publication Date: 2020-04-21
FRESHWATER FISHERIES RES CENT OF CHINESE ACAD OF FISHERY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the Yangtze finless porpoise lives in water, it is difficult to determine the genetic relationship between populations through observation and external characteristics, so a new, highly feasible technology is urgently needed to solve the problem of paternity identification of finless porpoise breeding populations

Method used

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  • A primer, kit and identification method for identifying the genetic relationship among individuals of the Yangtze finless porpoise
  • A primer, kit and identification method for identifying the genetic relationship among individuals of the Yangtze finless porpoise
  • A primer, kit and identification method for identifying the genetic relationship among individuals of the Yangtze finless porpoise

Examples

Experimental program
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Effect test

Embodiment 1

[0021] 1. Use the magnetic bead enrichment method to develop microsatellite markers for Yangtze finless porpoise, and select primers with high polymorphism and good amplification efficiency from multiple microsatellite markers by PCR amplification. Select 20 pairs of microsatellite primers, label different primers with FAM and HEX fluorescence, and detect two sites at the same time according to the fluorescence color. The primer sequences are shown in the table below.

[0022]

[0023] 2. Blood was collected from seven Yangtze finless porpoises, whose individual numbers were 1, 2, 3, 4, 5, 6, and 7. Among them, No. 3 is a mature female finless porpoise (assumed female parent), No. 4 and 5 are mature male finless porpoises (assumed male parent), and No. 1, 2, 6, and 7 are offspring bred by hypothetical parents. Take the blood of the above seven Yangtze finless porpoises, put them into a 1.5mL centrifuge tube, add 150μL of alkaline lysis solution (0.25mMNaOH, 0.25mM EDTA), he...

Embodiment 2

[0028] 1. Use the magnetic bead enrichment method to develop microsatellite markers for Yangtze finless porpoise, and select primers with high polymorphism and good amplification efficiency from multiple microsatellite markers by PCR amplification. 20 pairs of microsatellite primers were selected, and different primers were labeled with FAM and HEX fluorescence respectively, and two sites were simultaneously detected according to the fluorescent colors. The primer sequences were the same as in Example 1.

[0029] 2. Blood was collected from seven Yangtze finless porpoises, whose individual numbers were 1, 2, 3, 4, 5, 6, and 7. Among them, No. 3 is a mature female finless porpoise (assumed female parent), No. 4 and 5 are mature male finless porpoises (assumed male parent), and No. 1, 2, 6, and 7 are offspring bred by hypothetical parents. Take the blood of the above seven Yangtze finless porpoises, put them into a 1.5mL centrifuge tube, add 150μL of alkaline lysis solution (0.2...

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Abstract

The invention provides a primer, a kit and a method for identifying inter-individual half-sibling, full-sibling and parental relationships of Neophocaena asiaeorientalis asiaeorientalis. The primer comprises 20 pairs of microsatellite primers for amplification. The method is capable of identifying the inter-individual half-sibling, full-sibling and parental relationships of the Neophocaena asiaeorientalis asiaeorientalis rapidly and accurately and is widely applied to genetic resource conservation of the Neophocaena asiaeorientalis asiaeorientalis. But previous microsatellite studies are mostly used for identifying genetic structures and paternity of Neophocaena asiaeorientalis asiaeorientalis species and does not involve identification of the inter-individual half-sibling, full-sibling and parental relationships.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and in particular relates to a primer, a kit and an identification method for identifying half-siblings, full-siblings and parental kinship among Yangtze finless porpoise individuals. Background technique [0002] The Yangtze River finless porpoise (Neophocaena phocaenoides asiaeorientalis) is a small toothed whale listed as a national second-class protected animal. It is mainly distributed in the middle and lower reaches of the Yangtze River, Dongting Lake and Poyang Lake and their main tributaries. It is a relatively independent freshwater subspecies. According to the results of the international joint investigation of the Yangtze River porpoise at the end of 2006, the number was about 1,800 at that time, and it continued to decline at a rate of at least 5% per year. Since the ecological environment of the Yangtze River is unlikely to improve significantly in a relatively short period...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/156
Inventor 唐永凯徐跑俞菊华李建林刘凯李红霞于凡
Owner FRESHWATER FISHERIES RES CENT OF CHINESE ACAD OF FISHERY SCI
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