GSM3892121: RiboMethSeq shCtrl rep1; Homo sapiens; OTHER
Total RNA was extracted with Omega Total RNA Kit I (Omega) and quantified.Generally, 200 ng RNA was fragmented in a bicarbonate buffer (50 mM Sodium bicarbonate, pH 9.2) at 95 °C for 6 min. After purification, the RNA samples were 3'-end dephosphorylated using 5 U of Antarctic phosphatase (NEB) for 30 min at 37 °C. The 5'-end phosphorylation was subsequently performed using T4 PNK (NEB) at 37 °C for 1 hr. The RNA samples were purified with RNeasy MinElute Cleanup kit (Qiagen). The library construction was conducted using NEBNext Small RNA Library kit (NEB) following the manufacturer's instructions. Multiplexed libraries were sequenced with Illumina Hiseq4000 instrument in a 50 bp single-end read mode (Illumina).
GEO Accession
GSM3892121