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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

PCR-based genotyping of SNP markers in sheep

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Author(s):
Meo Niciura, Simone Cristina [1] ; Cruvinel, Giovanna Gabrielle [2] ; Moraes, Caroline Valerio [3] ; Bressani, Flavia Aline [1] ; Malago Junior, Wilson [1] ; Benavides, Magda Vieira [4] ; Souza de Chagas, Ana Carolina [1]
Total Authors: 7
Affiliation:
[1] Embrapa Pecuaria Sudeste, Rodovia Washington Luiz, Km 234, POB 339, BR-13560970 Sao Carlos, SP - Brazil
[2] Ctr Univ Cent Paulista, Rua Miguel Petroni 5111, BR-13563470 Sao Carlos, SP - Brazil
[3] Univ Fed Sao Carlos, Rodovia Washington Luiz, Km 235, BR-13566905 Sao Carlos, SP - Brazil
[4] Embrapa Pecuaria Sul, Rodovia BR-153, Km 632, 9 Vila Ind, BR-96401970 Bage, RS - Brazil
Total Affiliations: 4
Document type: Journal article
Source: MOLECULAR BIOLOGY REPORTS; v. 45, n. 4, p. 651-656, AUG 2018.
Web of Science Citations: 0
Abstract

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. (AU)

FAPESP's process: 17/01626-1 - Genetic and immune responses characterization associated with the phenotype of parasite resistance in a sheep flock of Morada Nova
Grantee:Ana Carolina de Souza Chagas
Support type: Regular Research Grants