GSM7866515: Marcela 24; Bos taurus; RNA-Seq
First, the CCs were removed by denudation and only oocytes with the presence of the first polar body (matured) were used in this analysis. cDNA synthesis and amplification from denuded oocytes were performed using the SMART-Seq HT Kit (Takara Bio Inc) and pools of 5 oocytes (~0.1 ng of total RNA) per group per replicate (3 replicates), following the manufacturer's recommendation. Next, cDNA was purified using AMPure XP Beads (Beckman Coulter), and cDNA concentration and profile were determined using Qubit dsDNA High Sensitivity (ThermoFisher Scientific) and Bioanalyzer High Sensitivity DNA Kit (Agilent), respectively. Libraries were prepared using the Nextera XT DNA Library Prep (Illumina) as recommended by the SMART-Seq HT kit. Libraries were assessed using Qubit and Bioanalyzer, before sequencing on the NextSeq (Illumina) considering 100 bp paired-end reads.