GSM9136144: Control-2; Homo sapiens; RNA-Seq
HeLa cells were lysed using IP lysis buffer (150 mM NaCl, 10 mM Tris-HCl pH 7.4, 1% NP-40, protease and RNase inhibitors). The lysates was then subjected to ultrasonic fragmentation at high power, 30 seconds on/30 seconds off for 10 cycles at 4°C. Cell lysates were incubated with antibody-conjugated magnetic beads overnight at 4°C. After immunoprecipitation, RNA was extracted using TRIzol reagent (Invitrogen) according to the manufacturer's instructions. RNA fragments were end-repaired and ligated with 3' and 5' adapters using the Small RNA-seq Library Preparation Kit for Illumina V3 (ABclonal, RK20312) and indexed using the Small RNA Adapter Module 96 Index (ABclonal, RK20358), following the manufacturer's instructions. cDNA was synthesized and PCR-amplified to generate sequencing-ready libraries.