GSM9141019: C57_MUS_CTRL_RM2; Mus musculus; RNA-Seq
The total RNA was extracted and purified from tumor tissues using TRIzol reagent. Nanodrop ND-2000 (Thermo Scientific, USA) was used to detect the A260/A280 absorbance ratio of the RNA samples, and Agilent Bioanalyzer 4150 (Agilent Technologies, CA, USA) was used to determine the RIN value of RNA. Only RNAs that passed the quality control were used for library construction. According to the ABclonal mRNA-seq Lib Prep Kit (ABclonal, China) instructions, PE libraries were prepared. The mRNA was purified from 1 μg of total RNA using oligo(dT) magnetic beads, followed by mRNA fragmentation in ABclonal First Strand Synthesis Reaction Buffer. Subsequently, the first strand of cDNA was synthesised using random primers and reverse transcriptase (RNase H) using the mRNA fragments as templates, and then the second strand of cDNA was synthesised using DNA polymerase I, RNAseH, buffer and dNTPs. The synthesised double stranded cDNA fragments were ligated to the junction sequence and used for PCR amplification. PCR products were purified and library quality was assessed using an Agilent Bioanalyzer 4150. Finally, sequencing was performed using Illumina Novaseq 6000 / MGISEQ-T7 sequencing platform.