title: Illumina HiSeq 2500 sequencing; Effect of cytokine treatment on mouse T helper cells
design: Effect of cytokine treatment on mouse T helper cells
library_construction_protocol: Naive CD4+ T cells were extracted from JAXJ mouse spleens using Stem Cell Technologies naive T cell purification kit. Stimuli were applied for a quick screen an unknown concentration to look for preliminary responses. T cells were grown in IMDM on CD3/CD28 coated round bottom 96w plates for 5 days Cytokines and drugs were added to the media 24h after plating Cells were spun down and lysed in RLT plus buffer. RNA was then extracted using SPRI beads Smartseq2 was used to generate and amplify cDNA. Nextera XT was used to generate the libraries