title: GSM2850561: RNAseq_JE1; Mus musculus; RNA-Seq
library_construction_protocol: 48 hours post-aspiration, all groups were euthanized for isolation of lung CD4+ T cells by magnetic bead pre-enrichment and fluorescence-activated cell sorting. RNA and genomic DNA were obtained using the AllPrep DNA/RNA Micro Kit (Qiagen). cDNA was generated with the Nugen Ovation RNA-Seq System V2. Fragmentation was performed on a Covaris S2. Illumina-compatible adapter ligation and barcoding was followed by PCR amplification. A High Sensitivity chip on an Agilent Bioanalyzer 2100 was used to measure the size distribution and quality of amplified libraries. Library quantification was performed with the qPCR-based Kapa Library Quantification Kit or by Bioanalyzer. Equimolar concentrations of each library were pooled. Cluster generation and sequencing were performed on an Illumina HiSeq 2500 employing 100 x 100 paired end sequencing with the TruSeq Rapid PE Cluster Kit and TruSeq Rapid SBS Kit (200 cycles).