Project description:We present RNA-Seq data obtained from HEK293T cells and HEK293T cells with ALKBH5 and FTO demethylases double knockout (HEK293TΔALKBH5ΔFTO), showing m6A-dependent transcriptome alterations.
Project description:RNA-Seq procedure was used for analysis of expression in HEK293T cells. Cells were separately lysed with Trizol reagent (Invotrogen). Total RNA was extracted from cell lysed with Trizol reagent and PureLink RNA Micro Kit (Invitrogen) according to manufacture instruction. RNA quality was checked with BioAnalyser and RNA 6000 Nano Kit (Agilent). Poly(A)+ RNA was purified with Dynabeads® mRNA Purification Kit (Ambion). Illumina library was prepared from poly(A)+RNA with NEBNextNEBNext® Ultra™II RNA Library Prep Kit for Illumina® (NEB) according to manual. Sequencing was performed on HiSeq1500 with 50 bp read length. At least 10 millions of reads were generated for each sample.
Project description:RNA-seq experiment to study the role of Pat1b in HEK293T cells. HEK293T cells were treated twice with siRNA with Lipofectamine 2000 then harvested after 48hours. Total RNA was extracted with TriReagent. Ribo-Zero TruSeq stranded mRNA libraries were prepared for each sample and sequenced on Illumina NextSeq 500 Sequencing System providing around 100 million reads per sample (around 75 bp paired-end reads).