GSM2100604: M phase TAIL-seq; Homo sapiens; OTHER
Total RNAs were extracted by TRIzol (Invitrogen) For TAIL-seq, 90 μg of total RNA extracted by TRIzol (Life Technologies) was DNase-treated and purified (>200 nt) by RNeasy MinElute cleanup column (QIAGEN). rRNAs were depleted by using Ribo-Zero kit (Epicentre) and remaining RNAs were ligated to the biotinylated 3′ adaptor and partially digested by RNaseT1 (Ambion). Fragmented RNAs were pulled-down by streptavidin beads and eluted RNAs were phosphorylated at the 5′ end. RNAs were size fractionated by gel purification (500-1,000 nt), and purified RNAs were ligated to 5′ adaptor, reverse-transcribed and amplified by PCR. PCR products were purified using AMPure XP beads (Beckman), and cDNA libraries were sequenced by Illumina HiSeq 2500 (51 × 251 bp paired end run) with PhiX control library and spike-in mixture with various poly(A) length.
GEO
GSM2100604