GSM5966595
Preparation of total RNA-seq library was performed using SMART-Seq Stranded Kit (Clontech), according to manufacturers' instruction. In brief, 19 of siScramble injected or 23 of siTardbp injected ZP-free embryos were lysed in 1× Lysis Buffer containing RNase inhibitor (0.2 IU/µl, from SMART-Seq Stranded Kit, Clontech), directly. RNAs were sheared by heating at 85℃ for 8 minutes and used for reverse transcription with random hexamers and PCR amplification. Ribosomal fragments were depleted from each cDNA sample with scZapR and scR-Probes. Indexed total RNA-seq libraries were enriched by second PCR amplification then sequenced using HiSeq Ⅹ sequencer (Illumina, paired-end, 150 bp). Three biological replicates were generated for each sample.