Abstract
Despite transcriptional silencing in mature sperm and cytoplasmic expulsion of RNA during the final sperm maturation process, thousands of RNAs have been successfully identified in ejaculated sperm. Although most of RNAs' function is still unknown, it is suggested that sperm RNAs have a vital biological role in fertilization and post-fertilization events. Nevertheless, the lack of accurate RNA isolation techniques and the resultant good quality sperm RNA has hampered the exploration of sperm RNAs function. Additionally, small non-coding RNAs are found in extracellular fluids including seminal plasma. These small RNAs may participate in cell to cell communication or intracellular and extracellular message transmission. Developing precise protocols to extract RNA from sperm and seminal plasma is critical to elucidate sperm physiology and paternal contributions to fertilization and post-fertilization events. A detailed procedure consisting of semen collection, separation of sperm and seminal plasma, extracting RNA from sperm and seminal plasma, and determining the quantity and quality of RNA for boar semen is presented here. This efficient protocol can be extrapolated to isolate RNAs from sperm and seminal plasma across mammalian species.