title: GSM8683548: single_Bryostatin_50; Mus musculus; RNA-Seq
library_construction_protocol: 1x10^6 live (Annexin V-) CD8+ T cells were sorted from the 4 different culture conditions and immediately lysed with TRIzol LS reagent (Life Technologies) and stored at -80°C. RNA was extracted according to manufacturer's instructions and a bioanalyzer (Agilent) was used to determine the integrity of the extracted RNA. NEBNext Ultra II Directional RNA Library Prep Kit for Illumina" (NEB #E7760S/L). mRNA was isolated from total RNA using the oligo-dT magnetic beads. After fragmentation of the mRNA, a cDNA synthesis was performed. This was used for ligation with the sequencing adapters and PCR amplicification of the resulting product. The quality and yield after sample preparation was measured with the Fragment Analyzer (see appendices). The size of the resulting products was consistent with the expected size distribution (a broad peak between 300-500 bp).