title: GSM8761521: Embryo_TAK_10; Mus musculus; RNA-Seq
library_construction_protocol: Total RNA was extracted from control and Tak-progeny embryos, collected at day E13.5 (n= 6 for control and n=12 for Tak-progeny) by guanidinium thiocyanate-phenol-chloroform extraction and DNAse treatment. Briefly, RNA was extracted and purified using a TRIzol-based protocol followed by acidic phenol/chloroform purification and DNase treatment to ensure RNA integrity and purity. Samples were homogenized in TRIzol (Invitrogen), followed by chloroform extraction and centrifugation to isolate the aqueous phase. RNA precipitation was achieved with isopropanol, followed by ethanol washes and air drying. Pellets were resuspended in RNase-free water. To enhance RNA quality, an acidic phenol/chloroform purification was performed twice, including Na-acetate addition and vortexing, with phases separated by centrifugation. Precipitation steps used isopropanol and ethanol, with overnight incubation at -20°C to enhance RNA recovery. DNAse treatment (DNAse I at concentration of 2u/µg of RNA) was conducted at 37°C for 30 minutes to remove residual DNA, followed by a final acidic phenol/chloroform purification and ethanol precipitation. Final RNA pellets were air-dried and dissolved in RNase-free water, with incubation at 50°C for optimal resuspension, ensuring RNA suitable for downstream applications. The mRNA libraries were performed by BGI Genomics using Hieff NGS Ultima Dual-mode mRNA kit (#12308ES96, Yeasen).