Data/hora: 20/06/2024 02:17:30

Biblioteca(s): | Embrapa Pecuária Sudeste.
Data corrente: | 10/08/2018
Data da última atualização: | 24/09/2018
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.
ISSN: | 1573-4978
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
Registro original: | Embrapa Pecuária Sudeste (CPPSE)
Exemplares: | CPPSE - (24421-1) - UPC - AP - DD - PROCI-2018.00040 - NIC - 2018.00052 - DIS