GSM2849982: KB4_K256L_A; Mus musculus; RNA-Seq
RNA was extracted from the T-cells using the RNeasy kit (QIAGEN) following the manufacturer instructions and total RNA was quantified using the Qubit RNA Assay Kit (Life Tech) and RNAquality was determined on the Bioanalyzer using the RNA Pico Kit (Agilent). The NuGen Ovation Human RNA-Seq Multiplex system (NuGen, part 0341) Prep Kit, was used to target deletion of unwanted high abundance transcripts and ribosomal RNA. More than 100ng of total RNA was converted into each DNA library following the manufacturer's protocol without modification. Following library construction, DNA libraries were quantified using the Qubit High Sensitivity DNA Kit (Life Tech) and library size was determined using the Bioanalyzer High Sensitivity Chip Kit (Agilent). Finally, qPCR was carried-out on the libraries using the Universal Library Quantification Kit for Illumina (Kapa Biosystems) and run on the 7900HT Fast qPCR machine (ABI). Libraries passing quality control were diluted to 2nM in sterile water and then sequenced on the NextSeq500 (Illumina) at a final concentration of 12pM, following all manufacturer protocols.