{"title":"Direct single-nucleotide polymorphism genotyping from whole blood without DNA extraction.","authors":"Risako Furukawa, Teruaki Tozaki, Koki Kawate, Mio Kikuchi, Taichiro Ishige, Emiko Fukui, Hironaga Kakoi","doi":"10.1294/jes.37.35","DOIUrl":null,"url":null,"abstract":"<p><p>A subset of single-nucleotide polymorphisms (SNPs) are associated with various phenotypes, including diseases, coat colour, and athletic performance, and are widely used in medical and veterinary fields. In this study, we developed a novel method for direct SNP genotyping from whole blood without DNA extraction. Thoroughbred blood samples were diluted 100-fold with Milli-Q water and analysed using real-time polymerase chain reaction (PCR) with hydrolysis probes. Specificity and sensitivity were improved by increasing the annealing temperature and number of PCR cycles. Genotyping results for SNPs in <i>MSTN</i> and <i>LCORL</i> showed complete concordance with conventional real-time PCR using the extracted DNA. This method is simple, low-cost, highly versatile, and applicable to other genetic targets, such as <i>CDH13</i> and <i>MAOA</i>.</p>","PeriodicalId":35701,"journal":{"name":"Journal of Equine Science","volume":"37 1","pages":"35-40"},"PeriodicalIF":0.0000,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12995552/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Equine Science","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1294/jes.37.35","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2026/3/14 0:00:00","PubModel":"Epub","JCR":"Q3","JCRName":"Veterinary","Score":null,"Total":0}
引用次数: 0
Abstract
A subset of single-nucleotide polymorphisms (SNPs) are associated with various phenotypes, including diseases, coat colour, and athletic performance, and are widely used in medical and veterinary fields. In this study, we developed a novel method for direct SNP genotyping from whole blood without DNA extraction. Thoroughbred blood samples were diluted 100-fold with Milli-Q water and analysed using real-time polymerase chain reaction (PCR) with hydrolysis probes. Specificity and sensitivity were improved by increasing the annealing temperature and number of PCR cycles. Genotyping results for SNPs in MSTN and LCORL showed complete concordance with conventional real-time PCR using the extracted DNA. This method is simple, low-cost, highly versatile, and applicable to other genetic targets, such as CDH13 and MAOA.
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