GSM8162197: SN_rep1; Mus musculus; RNA-Seq
The zona pellucidae of oocytes were removed with 0.5% pronase E (Sigma). 15 oocytes per sample (3 samples for each group) were lysed and subjected to first strand synthesis. The transcriptome analysis of GV oocytes was conducted via Smart-seq2, following our previously described methodology(49). 15 oocytes were lysed in 0.2% Triton X-100 containing 80 U RNase Inhibitor. First-strand cDNA synthesis involved the Oligo(dT) VN primer, followed by the 5'TS oligo primer. Subsequently, cDNA amplification was carried out using an ISPCR primer in an 18-cycle PCR and the products underwent purification via VAHTS DNA Clean Beads (Vazyme, N411). Quality control for the amplification products involved measuring cDNA concentration with Qubit 4 and assessing cDNA fragment distribution with Qsep 400. Following this, sequencing libraries were prepared with the TruePrep DNA Library Prep Kit for Illumina (Vazyme, TD503).