title: Optimized methods for extracting circulating small RNAs from long-term stored equine samples.
abstract: Circulating miRNAs in body fluids, particularly serum, are promising candidates for future routine biomarker profiling in various pathologic conditions in human and veterinary medicine. However, reliable standardized methods for miRNA extraction from equine serum and fresh or archived whole blood are sorely lacking. We systematically compared various miRNA extraction methods from serum and whole blood after short and long-term storage without addition of RNA stabilizing additives prior to freezing. Time of storage at room temperature prior to freezing did not affect miRNA quality in serum. Furthermore, we showed that miRNA of NGS-sufficient quality can be recovered from blood samples after >10 years of storage at -80 °C. This allows retrospective analyses of miRNAs from archived samples.
authors.affiliation: Department of Clinical Veterinary Medicine, Swiss Institute of Equine Medicine, Vetsuisse Faculty, University of Bern and Agroscope, Länggassstrasse 124, 3012, Bern, Switzerland.
authors.forename: Lucia
authors.initials: L
authors.lastname: Unger
authors.affiliation: Department of Clinical Veterinary Medicine, Swiss Institute of Equine Medicine, Vetsuisse Faculty, University of Bern and Agroscope, Länggassstrasse 124, 3012, Bern, Switzerland.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Fouché
authors.affiliation: Department of Clinical Research and Veterinary Public Health, Institute of Genetics, Vetsuisse Faculty, University of Bern, Bremgartenstrasse 109A, 3012, Bern, Switzerland.
authors.forename: Tosso
authors.initials: T
authors.lastname: Leeb
authors.affiliation: Department of Clinical Veterinary Medicine, Swiss Institute of Equine Medicine, Vetsuisse Faculty, University of Bern and Agroscope, Länggassstrasse 124, 3012, Bern, Switzerland.
authors.forename: Vincent
authors.initials: V
authors.lastname: Gerber
authors.affiliation: Department of Clinical Veterinary Medicine, Swiss Institute of Equine Medicine, Vetsuisse Faculty, University of Bern and Agroscope, Länggassstrasse 124, 3012, Bern, Switzerland. alicja.pacholewska@vetsuisse.unibe.ch.|Department of Clinical Research and Veterinary Public Health, Institute of Genetics, Vetsuisse Faculty, University of Bern, Bremgartenstrasse 109A, 3012, Bern, Switzerland. alicja.pacholewska@vetsuisse.unibe.ch.
authors.forename: Alicja
authors.initials: A
authors.lastname: Pacholewska
mesh_terms: D000818:Animals; D001774:Blood Chemical Analysis / Q000379:methods*; D005615:Freezing; D006736:Horses; D035683:MicroRNAs / Q000097:blood* / Q000302:isolation & purification*; D013048:Specimen Handling
chemical_list: D035683:MicroRNAs
keywords: EDTA; Horse; RNA extraction; Serum; Small RNA; miRNA; microRNA