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m6A-Seq

BL Boyang Liu
JZ Jian Zhou
CW Chenyang Wang
YC Yajie Chi
QW Quantang Wei
ZF Zhao Fu
CL Changlin Lian
QH Qiongzhen Huang
CL Chenxin Liao
ZY Zhao Yang
HZ Huijun Zeng
NX Ningbo Xu
HG Hongbo Guo
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m6A-IP and library preparation were carried out according to the reported procedure45. In brief, poly-A-purified RNA was fragmented with Ambion RNA Fragmentation reagent (Ambion, Carlsbad, CA, USA), and incubated with m6A primary antibody at 4°C for 2 h. The mixture was immunoprecipitated through incubation with Protein A beads (Thermo Fisher, MA, USA) at 4°C for 2 h. Followed by washing for three times, the Captured RNA was eluted with m6A nucleotide solution and purified with RNA Clean and Concentrator Kit (Zymo, LA, USA). Sequencing was performed on Illumina HiSeq 2000 according to the manufacturer’s recommendations.

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