GSM8840993: MII oocytes vivo 1; Bos taurus; RNA-Seq
DNA and RNA from oocyte were separated using the G&T protocol (Angermueller et al. 2016). The magnetic beads (MyOne C1, Life Technologies) were washed and then annealed to oligo dTs. These were used to capture polyadenylated mRNA from individual cell lysate. The supernatant containing the DNA was transferred to a new tube and the beads washed three times in 1xFSS buffer (Superscript II, Invitrogen), 10 mM DTT, 0.005% Tween-20 (Sigma) and 0.4 U/μl of RNAsin (Promega) to remove all DNA residue. Each washing solution was added to the DNA tube to maximise recovery. Total RNA was extracted from seven cumulus cells samples (four from in vivo matured, three from in vitro matured COCs) using RNeasy Micro Kit (Qiagen) according to the manufacturer's instruction. The RNA samples underwent quality and concentration assessments using an RNA 6000 Pico Chip (Agilent Technologies, Carlsbad, CA, USA). ScRNA-seq and scBSseq