基于微卫星多重PCR技术的特克塞尔×哈萨克杂交羊亲子鉴定

doi:10.16431/j.cnki.1671-7236.2020.07.022

Abstract

Abstract: The purpose of this study was to establish a set of identification system for parent-child relationship of Texel×Kazakh sheep.In the experiment,11 microsatellite marker sites were selected for combined amplification.By optimizing the primer concentration,annealing temperature,and reaction system of the combination of microsatellite marker sites,4 sets of multiplex PCR systems were established.Capillary electrophoresis was performed after multiplex PCR to perform genotyping to identify.The genotyping results were read by PROSize 3.0 software,and the gene-tic diversity of the population was analyzed by Cervus 3.0 software.The parent-child relationship between Texel×Kazakh progressive F2 generation (Texel×Kazakh progressive F2) and cross F2 generation (Texel×Kazakh cross F2).The results showed that the alleles of the Texel×Kazakh progressive F2 and cross F2 were 180 and 140,the average numbers of alleles were 16.364 and 12.727,the average observed heterozygosity were 0.533 and 0.544,the average expected heterozygosity were 0.807 and 0.831,and the average polymorphism information content were 0.783 and 0.803.Parent-child relationship identification was performed at 11 microsatellite marker sites of 70 candidate parents and 64 candidate progenies of the Texel×Kazakh progressive F2 and cross F2 two breeds.The results showed that:When the parental genotype was unknown,the combined exclusion probability (CE-1P) was 0.9995 and 0.9997;when the parental genotype was known,the combined exclusion probability (CE-2P) was all reached 1.0000;And the combined exclusion probability of both parents reached 1.0000.It indicates that the 11 selected microsatellite marker sites had high polymorphism and high probability of exclusion,which was suitable for genetic analysis and paternity identification of individuals.The parent-child identification system of Texel×Kazakh sheep established by microsatellite markers provided a theoretical basis for analyzing genetic diversity of sheep populations and assisting breeding work.

Key words: Texel×Kazakh hybrid sheep; microsatellite; multiplex PCR; paternity test

CLC Number:

S813.22

LI Bin, REN Tingting, QIU Meiyu, LI Zhiqiang, YAN Xiaofei, HAN Bing, LIU Mingjun, HE Sangang, YIMING·Sulaiman. Paternity Identification of Texel×Kazakh Sheep Based on Microsatellite Multiple PCR Technology[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(7): 2171-2180.

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Paternity Identification of Texel×Kazakh Sheep Based on Microsatellite Multiple PCR Technology

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