RNAseq of DM1-B myotubes treated with 7.8nM MPC04
100ng of total RNA from DM1-B MR (NJ5-21) myotubes treated with 7.8nM MPC04 was used to generate RNA-sequencing libraries. RNA integrity numbers (RIN) were measured via capillary gel electrophoresis using a fragment analyzer (Agilent) with the Agilent DNF-471 kit. RNA sample was prepared for sequencing using the NEBNext Ultra II Direction RNA library Prep Kit for Illumina with ribosomal RNA depletion via the NEBNext rRNA Depletion Kit. The following changes were made to the manufacturers protocol: 40x adaptor dilutions were made, bead incubations were all performed at room temperature, and 15 cycles of library amplification were used. Libraries were quantified for quality and yield using the NEB-Next Library Quant Kit and capillary gel electrophoresis using the Agilent DNF-474 kit. Libraries were pooled in equimolar concentrations and sequenced using paired end 100 base pair sequencing on the Illumina NextSeq 2000 sequencer at the University at Albany.