Profiling of MicroRNAs in the Biofluids of Livestock Species
The value of circulating microRNAs (miRNAs) as noninvasive biomarkers of human disease has been extensively demonstrated. Significant potential also exists in other species, particularly in relation to control of veterinary diseases and selection/monitoring of production traits in livestock. Although robust protocols have been developed for miRNA profiling of human biofluids, significant optimization may be required before these can be applied to other species. In this chapter, we describe protocols for small-RNA sequencing and RT-qPCR analyses of plasma samples from livestock species. In addition, we provide brief data analysis protocols for small-RNA sequencing and RT-qPCR data. Finally, we highlight important considerations for these protocols such as low RNA yield, platform-specific biases, and optimal normalization approaches.
Key wordsCow miRNA microRNA Biomarker Plasma Follicular fluid Sequencing
We would like to thank Bushra Mohammed, Stephanie Schauer, and Sadanand Sontakke for assistance with developing protocols. This work was funded by Zoetis Inc. and BBSRC.
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