RNA seq in CD8T cells

Purpose: we evaluated the T cell mediated anti tumor immune response under asparagine restriction Methods: Total RNA was isolated according to the RNeasy Mini Kit (Qiagen) and treated with DNase I. RNA quality was assessed using the Agilent 2100 Bioanalyzer and RNA Nano Chip kit (Agilent Technologies, CA). RNA-seq libraries were generated using TruSeq Stranded Total RNA with Ribo-Zero Globin Complete kit (Illumina, CA). Approximately 60–80 million paired-end 150 bp reads were generated per sample using Illumina HiSeq 4000 platform Results: We performed gene expression profiling analysis using data obtained from RNA-seq of T cells cultured in presence and absence of asparagine at 3 different time points and found that asparagine restriction induces biphasic response in T cell. mRNA profiles of CD8 T cells were generated by deep sequencing, using Illumina HiSeq 4000.