GSM5391161: small_RNA_NT.Bl.4; Bos taurus; ncRNA-Seq
RNA was isolated based on size using the RNA Clean & ConcentratorTM 5 kit (Zymo) from three pools each of oocytes, 2-cell stage embryos, 8-cell stage embryos, morula stage embryos, or blastocyst stage embryos according to the manufacturer's protocol for purification of small RNA (<200 nt) and large RNA (>200 nt) as separate fractions. Small RNA sequencing was performed on the Ion Proton Sequencer by using the Ion Total RNA-seq kit v2 (Thermo Fisher Scientific) according to manufacturer's procedure for small RNA library preparation with no deviations from the specified protocol. By using an RNA isolation protocol that yielded a specific fraction of small RNA, an enrichment step was not needed.  Sample volumes were reduced to 3 l by vacuum centrifugation, and the entire sample was used to prepare the small RNA library for sequencing.  The cDNA sample was then processed on the Agilent Bioanalyzer (Agilent, Santa Clara, California) to ensure the presence of small RNA bound to a barcode (86 to 106 nt).