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Provedor de dados:  Repositório Alice
País:  Brazil
Título:  PCR-based genotyping of SNP markers in sheep.
Autores:  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.
Data:  2018-08-10
Ano:  2018
Palavras-chave:  Tetra primer ARMS PCR
PCR RFLP
Resistência nematódeo gastrintestinal
Marcador Molecular
Ovino
Genome
Resumo:  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.

bitstream/item/181282/1/PCR-based.pdf
Tipo:  Artigo em periódico indexado (ALICE)
Idioma:  Inglês
Identificador:  1573-4978

24421

http://www.alice.cnptia.embrapa.br/alice/handle/doc/1094070

https://doi.org/10.1007/s11033-018-4206-8
Editor:  Molecular Biology Reports, v.45, n.4, p.651-654, aug. 2018.
Relação:  Embrapa Pecuária Sudeste - Artigo em periódico indexado (ALICE)
Formato:  p.651-654
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