02277naa a2200277 a 450000100080000000500110000800800410001902400510006010000220011124500730013326000090020652014620021565000110167765000230168865000100171165300130172165300440173465300260177870000200180470000180182470000230184270000220186570000210188770000240190877300670193221046942019-01-23       2018    bl ---       0--  u    #d7 ahttps://doi.org/10.1007/s11033-018-4206-82DOI1 aNICIURA, S. C. M.  aPCR-based genotyping of SNP markers in sheep.h[electronic resource]  c2018  aSingle 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.  aGenome  aMarcador Molecular  aOvino  aPCR RFLP  aResistência nematódeo gastrintestinal  aTetra primer ARMS PCR1 aCRUVINEL, G. G.1 aMORAES, C. V.1 aDONATONI, F. A. B.1 aMALAGO JUNIOR, W.1 aBENAVIDES, M. V.1 aCHAGAS, A. C. de S.  tMolecular Biology Reportsgv. 45, n. 4, p. 651-654, Aug. 2018.