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利用新一代测序技术进行高通量且经济高效的鸡基因分型

High-throughput and Cost-effective Chicken Genotyping Using Next-Generation Sequencing.

作者信息

Pértille Fábio, Guerrero-Bosagna Carlos, Silva Vinicius Henrique da, Boschiero Clarissa, Nunes José de Ribamar da Silva, Ledur Mônica Corrêa, Jensen Per, Coutinho Luiz Lehmann

机构信息

Animal Biotechnology Laboratory, Animal Science and Pastures Department, University of São Paulo (USP)/Luiz de Queiroz College of Agriculture (ESALQ), Piracicaba, São Paulo, Brazil.

IFM Biology, AVIAN Behavioural Genomics and Physiology Group, Linköping University, Linköping, Sweden.

出版信息

Sci Rep. 2016 May 25;6:26929. doi: 10.1038/srep26929.

Abstract

Chicken genotyping is becoming common practice in conventional animal breeding improvement. Despite the power of high-throughput methods for genotyping, their high cost limits large scale use in animal breeding and selection. In the present paper we optimized the CornellGBS, an efficient and cost-effective genotyping by sequence approach developed in plants, for its application in chickens. Here we describe the successful genotyping of a large number of chickens (462) using CornellGBS approach. Genomic DNA was cleaved with the PstI enzyme, ligated to adapters with barcodes identifying individual animals, and then sequenced on Illumina platform. After filtering parameters were applied, 134,528 SNPs were identified in our experimental population of chickens. Of these SNPs, 67,096 had a minimum taxon call rate of 90% and were considered 'unique tags'. Interestingly, 20.7% of these unique tags have not been previously reported in the dbSNP. Moreover, 92.6% of these SNPs were concordant with a previous Whole Chicken-genome re-sequencing dataset used for validation purposes. The application of CornellGBS in chickens showed high performance to infer SNPs, particularly in exonic regions and microchromosomes. This approach represents a cost-effective (~US$50/sample) and powerful alternative to current genotyping methods, which has the potential to improve whole-genome selection (WGS), and genome-wide association studies (GWAS) in chicken production.

摘要

鸡的基因分型在传统动物育种改良中已成为常见做法。尽管高通量基因分型方法功能强大,但其高昂成本限制了在动物育种和选择中的大规模应用。在本文中,我们对康奈尔简化基因组测序法(CornellGBS)进行了优化,该方法是一种在植物中开发的高效且经济的测序基因分型方法,以使其适用于鸡。在此,我们描述了使用康奈尔简化基因组测序法对大量鸡(462只)成功进行基因分型的情况。基因组DNA用PstI酶切割,连接到带有识别个体动物条形码的接头,然后在Illumina平台上进行测序。应用过滤参数后,在我们的实验鸡群体中鉴定出134,528个单核苷酸多态性(SNP)。在这些SNP中,67,096个具有至少90%的分类群检出率,被视为“独特标签”。有趣的是,这些独特标签中有20.7%此前未在单核苷酸多态性数据库(dbSNP)中报道。此外,这些SNP中有92.6%与先前用于验证目的的全鸡基因组重测序数据集一致。康奈尔简化基因组测序法在鸡中的应用显示出在推断SNP方面的高性能,特别是在外显子区域和微染色体中。这种方法是一种经济高效的(约50美元/样本)且强大的当前基因分型方法的替代方法,有潜力改善鸡生产中的全基因组选择(WGS)和全基因组关联研究(GWAS)。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d93b/4879531/c20fa2872510/srep26929-f1.jpg

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