title: GSM5107448: Sperm miRNA; Bos taurus; miRNA-Seq
library_construction_protocol: Sperm were filtered from the diced cauda epididymis, and extracted by trituration at room temperature. Spermatozoa from whole pieces were removed and collected using the swim‐up method in BO solution pre‐warmed to 37°C. After incubating the sample for 30 min at 37°C with the tube slanted at an approximate angle of 45°, the motile overlying fraction (1 ml liquid) was pipetted into a clean tube containing 1 ml of HEPES‐buffered Tyrode's lactate sperm medium (100 mM NaCl, 25 mM NaHCO3, 10 mM HEPES, 0.3 mM NaH2PO4, 3.11 mM KCl, 0.035 mM sodium lactate, 2 mM CaCl2·2 H2O, 4 mM MgCl2·6 H2O [pH 7.3]) (Parrish et al., 1995). This mixture was centrifuged at 800g for 5 min at 30°C. The pelleted sperm were evaluated (see below), and then frozen at −80°C until RNA isolation. RNA libaries were prepared for sequencing using standard Illumina protocols