Sequencing of mRNA in AML cells upon ALKBH5 knockdown

To identify the potential mRNA targets of ALKBH5, we conducted RNA-seq of mRNA samples enriched from AML cells with ALKBH5 knockdown Overall design: AML cells (herein is NOMO1 and MOLM13) were transduced with lentivirus expressing shRNAs, then selected with puromycin for 4 days. Polyadenylated RNAs (poly(A) RNAs) were extracted with FastTrack MAG Maxi mRNA isolation kit (Life technology) and randomly fragmented with RNA fragmentation reagents (Ambion). Final library preparation was constructed with TruSeq Stranded mRNA Sample Prep Kit (Illumina) and the library was quantified by BioAnalyzer High Sensitivity DNA chip. The RNA-seq was conducted on the Illumina HiSeq 2500.