ICA-H-01-1
Total cellular RNA was isolated from PBMCs with the RNeasy Mini kit (QIAGEN). Multiplexed RNA libraries were prepared with the Truseq RNA sample prep kit (Illumina). In brief, poly(A)-containing mRNAs were captured with oligodT beads, fragmented, reverse-transcribed, and the cDNA was ligated to Illumina adapters containing indexing barcodes. Libraries were quantified with KAPA Library Quant kits (KAPA Biosystems), and run on a HiSeq 2000 Sequencing System (Illumina) to produce 150 bp single-end reads. Reads were then mapped with STAR v2.5.3a. Sashimi plots were generated with IGV (Broad Institute), using a minimum threshold of 20 reads for a splice event to be taken into account.