Next Generation Sequencing Facilitates Quantitative Analysis of Msx1-overexpressed and Control C2C12 Cells Transcriptomes

Purpose: RNA-seq analysis to evaluate the impact of Msx1-overexpression on the transcriptome of C2C12 cells. Methods: Total RNA was isolated with TRIzol from Msx1-overexpressing C2C12 cells (n = 3) or control cells (n = 3). cDNA sequencing libraries were prepared with an NEBNext® Ultra™ RNA Library Prep Kit for Illumina. The RNA-Seq FASTQ raw data were trimmed to remove low-quality reads and adapters using Trimmomatic. The trimmed reads were aligned to the mouse reference genome UCSC GRCm38/mm10 with HISAT2. Gene and transcript quantification was performed using StringTie. The results of the mapping of the RNA-seq reads, transcript assembly and abundance estimation were reported as fragments per kilobase of exon per million fragments mapped (FPKM). mRNA profiles of Msx1-overexpressing C2C12 cells (n = 3) or control cells (n = 3).