GSM8223622: EpiLC, primed pluripotency, WT, rep1; Mus musculus; RNA-Seq
Total RNA was isolated from cells using the RNeasy Plus Mini Kit (QIAGEN 74136). RNA was treated with DNAse using the TURBO DNA-free kit (Invitrogen™ AM1907) prior to prepping libraries. RNA quality was assessed by electrophoresis on a 1% agarose gel and detecting 28S and 18S rRNA bands with ethidium bromide staining (0.6µg/mL). Illumina Stranded Total RNA Prep with Ribo-Zero Plus kit (Illumina 20040525) was used following the manufacturer's protocol to prep libraries with a starting RNA input of 500ng. Libraries were amplified using 11 cycles on the thermocycler. Each library was uniquely indexed with IDT for Illumina–DNA/RNA UD Indexes (Illumina 20026121), then pooled together in equimolar amounts. Library concentration was measured using the Qubit dsDNA HS assay kit (Invitrogen Q32851) and Qubit 4 fluorometer. Library size was assayed using the Agilent Bioanalyzer 2100 with the High sensitivity DNA kit.