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.Electronic supplementary materialThe online version of this article (doi:10.1186/s13028-016-0224-5) contains supplementary material, which is available to authorized users.
Highlights
The aim of our study was to compare different methods for the isolation of micro RNA (miRNA) from equine serum, fresh and archived ethylenediamine tetraacetic acid (EDTA) blood samples
The knowledge about micro RNA (miRNA) in physiologic and pathologic conditions is still very limited restricted to certain diseases only, and warrants further investigation in this species [3,4,5,6]
In contrast to mRNA, miRNA molecules are surprisingly stable—even in tissue samples of compromised quality [7]—which supports the possibility of routine use of miRNA fingerprints as biomarkers in clinical practice
Summary
The aim of our study was to compare different methods for the isolation of miRNAs from equine serum, fresh and archived EDTA blood samples. For miRNA extraction from serum samples we assessed how RNA yield is affected by the performance of three column-based and one column-free method, two different lysis reagents, and the effects of storage time at room temperature (RT), serum input volume, performing a second phenol extraction, and a second column elution.
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
