GSM3408806: ABA_0h_poly(A)-; Arabidopsis thaliana; RNA-Seq
Total RNA was extracted with TRIzol (Invitrogen) and treated with DNase I (Ambion). Polyadenylated [poly(A)+] RNAs were isolated from total RNA through two rounds of purification with oligo(dT) beads (QIAGEN). The poly(A)+ RNA depleted fraction from the first round of purification was collected as crude non-polyadenylated [(poly(A)-] RNA sample, which was subjected to another two rounds of treatment with oligo(dT) beads. Ribosomal RNAs were removed by two rounds of treatment with the Ribo-minus kit (Invitrogen). Libraries for RNA-seq were generated using NEXTflex Directional RNA-Seq Kit (dUTP based)  according to the manufacturer's instructions.