GSM6591852: A549 flag CTRL Rep3; Homo sapiens; RIP-Seq
cells were harvested and lysed in RIP lysis buffer (20mM Tris-HCl, pH7.4, 10mM MgCl2, and 0.1M NaCl, 10% glycerol, 0.5% NP-40, 0.5% Triton X-100, phosphatase inhibitors, protease inhibitors cocktail and RNase inhibitor) on ice for 5 min, followed by homogenizing for additional 20 min to release the nuclear components. Equivalent amount of RNA was mixed with spike-in (Invitrogen #4456740) to generate RIP-hnRNP M libraries using VAHTS Universal V6 RNA-seq Library Prep Kit (Vazyme #NRM604-01), according to the manufacture instruction.