Total RNAs were prepared using Trizol Reagent and treatment with DNase I. 20μg of total RNAs were separated into polyadenylated and nonpolyadenylated fraction by oligo(dT) magnetic beads (Invitrogen). Ribosomal RNAs were further depleted in nonpolyadenylated fraction by using RiboMinus kit (Invitrogen) to get the poly(A)-/ribo- RNA population. Poly(A)-/ribo- RNAs were further incubated with 5U of RNase R (Epicentre) for 3h at 37℃ and then were subjected to Trizol extraction (Invitrogen). An aliquot of poly(A)-/ribo- enriched and RNase R digested RNA was used for library preparation with Illumina TruSeq Stranded Total RNA HT Sample Prep Kit (P/N 15031048), and then subjected to 1x100 deep sequencing with Illumina HiSeq 2000
GEO Accession
GSM1164885