Transcriptome sequencing of murine myeloid leukemia genome
Experimental procedures. Whole Transcriptome RNA-seq (wtRNA-seq) was performed using total RNA isolated from flash frozen spleen specimens using mirVana™ miRNA Isolation Kit. 5 μg of total RNA plus ERCC RNA Spike-In was used for selective ribosomal RNA (rRNA) depletion using the RiboMinus™ Eukaryote System v2. 500 ng of rRNA-depleted total RNA was used for whole transcriptome library construction according to the Ion Total RNA-Seq Kit for the AB Library Builder™ System protocol for barcoded libraries, Ion Sphere™ Particle (ISP) loading using Ion OneTouch™ 2 System and sequencing on the Ion Proton™ platform with PIv2 semiconductor wafer chips per manufacturers recommended instructions. Each RNA-seq library was sequenced individually on a PIv2 chip to achieve >4 GB (~70 million reads) per specimen. After quality control checks, raw sequencing read files were generated by Ion Torrent Suite.
Houston Methodist Research Institute
D0005_SP_wtRNA-Seq_T