GSM9414485: RNA_13hpi_control-overexpressing_rep2; Mus musculus; RNA-Seq
Embryos in this study were derived from  In Vitro fertilization (IVF).Oocytes and embryos were collected at indicated time point.For RNA-seq, the collected samples were immediately transferred in lysis buffer (0.2% Triton X-100 and 2 U/ul RNase inhibitor) and then frozen at -80℃. For Ribo-seq, the collected samples were were lysed in iced lysis buffer (20 mM Tris-HCl pH 7.4, 150 mM NaCl, 5 mM MgCl2, 1 mM DTT, 100 μg/ml CHX, 1% Triton X-100; 25 U/ml Turbo DNase)  and then frozen at -80℃. The RNA-seq libraries for the collected samples were generated following the Smart-seq2 methodology.Sequnecing libraries were generated with TruePrep DNA Library Prep Kit (Vazyme, TD502) according to the manufacturer's guidance. For Ribo-seq, the lysate was then treated with 1 ul RNaseI 100 U/μ and incubated at room temperature for 45 min with gentle mixing. The nuclease digestion was stopped by adding 10μl SUPERase·In. Subsequently, ribosome-protected fragments (RPFs) were separated by the conventional Ribo-seq. RPFs library construction was conducted using Ezra-seq approach as previously described.