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Biblioteca(s): |
Embrapa Pecuária Sul. |
Data corrente: |
23/01/2019 |
Data da última atualização: |
23/01/2019 |
Tipo da produção científica: |
Artigo em Periódico Indexado |
Autoria: |
NICIURA, S. C. M.; CRUVINEL, G. G.; MORAES, C. V.; DONATONI, F. A. B.; MALAGO JUNIOR, W.; BENAVIDES, M. V.; CHAGAS, A. C. de S. |
Afiliação: |
SIMONE CRISTINA MEO NICIURA, CPPSE; Giovanna Gabrielle Cruvinel, UNICEP; Caroline Valério Moraes, UFSCar; FLAVIA ALINE BRESSANI DONATONI, CPPSE; WILSON MALAGO JUNIOR, CPPSE; MAGDA VIEIRA BENAVIDES, CPPSUL; ANA CAROLINA DE SOUZA CHAGAS, CPPSE. |
Título: |
PCR-based genotyping of SNP markers in sheep. |
Ano de publicação: |
2018 |
Fonte/Imprenta: |
Molecular Biology Reports, v. 45, n. 4, p. 651-654, Aug. 2018. |
DOI: |
https://doi.org/10.1007/s11033-018-4206-8 |
Idioma: |
Inglês |
Conteúdo: |
Single nucleotide polymorphisms (SNPs) are the main type of variation in genome, enabling them to be associated with traits of economic importance in livestock. Genome-wide association studies (GWAS) have led to the discovery of SNPs associated with desirable traits in sheep. However, in these studies, SNPs are genotyped by high-throughput methods in genome scale, which are expensive and require sophisticated equipment and analysis methods. Therefore, the goal of this study was to develop a reliable, rapid, and inexpensive polymerase chain reaction (PCR)-based method to genotype a medium number of animals for a few candidate SNPs previously associated with desirable phenotypes in sheep by GWAS, using markers associated with gastrointestinal nematode resistance as a model. DNA extracted from white-blood cells of 150 sheep was submitted to PCR amplification followed by agarose gel electrophoresis and determination of banding pattern. Tetra-primer ARMS-PCR was successfully optimized after changes in annealing temperature; annealing and extension times; concentration of MgCl2 and DNA; ratios of inner, outer, forward and reverse primer; and addition of adjuvants, for genotyping the OAR2_14765360, OAR6_81718546, OAR11_62887032, and OAR12_69606944 SNPs in sheep. An extensive optimization of tetra-primer ARMS-PCR resulted in a suitable, simple, cost-effective PCR-based method of genotyping four SNP markers previously detected by chip arrays. |
Palavras-Chave: |
PCR RFLP; Resistência nematódeo gastrintestinal; Tetra primer ARMS PCR. |
Thesagro: |
Marcador Molecular; Ovino. |
Thesaurus Nal: |
Genome. |
Categoria do assunto: |
X Pesquisa, Tecnologia e Engenharia |
Marc: |
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Registro original: |
Embrapa Pecuária Sul (CPPSUL) |
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