GSM3504938: s4U-RNA rep2; Homo sapiens; RNA-Seq
3×10^6 HEK293T cells were seeded per 10-cm cell dish and grown for 24 h. For T > C conversion identification, 0 or 50 μM s4U was added to the medium and grown for another 24 h; For mRNA half-life calculation, 500 μM s4U was added to the medium and grown for 1 h; For PDP-mediated RNA dynamic analysis, PDP (0 or 4 or 8 μM) was added to the medium for 20 min prior to 1 h 500 μM s4U incubation. After that, RNA samples for library construction were prepared by total RNA extraction, polyadenylated RNA isolation, gDNA removal, RNA fragmentation and  acrylonitrile treatment. For T > C conversion identification, we named the RNA samples as Con-RNA (no s4U treatment, no acrylonitrile treatment), s4U-RNA (s4U treatment, no acrylonitrile treatment), Con-acrylonitrile-RNA (no s4U treatment, acrylonitrile treatment) and s4U-acrylonitrile-RNA (s4U treatment, acrylonitrile treatment), respectively. For mRNA half-life calculation, we named the RNA samples as halflife-RNA.  For PDP-mediated RNA dynamic analysis,  we named the RNA samples as PDP-0-RNA (no PDP treatment), PDP-4-RNA (4 μM PDP treatment) and PDP-8-RNA (8 μM PDP treatment), respectively. Standard RNA seq libraries were prepared using NEBNext® Ultra™ II Directional RNA Library Prep Kit for Illumina® (NEB) according to manufacturer's instructions. Sequencing was performed on Illumina Illumina HiSeq X Ten.
GEO Accession
GSM3504938