GSM8332651: RNA_DE.L_C57B6_5.1; Mus musculus; RNA-Seq
Phase separation in cells lysed in TRIzol was induced with 50% isopropanol and 0.5% 2-mercaptoethanol and RNA was extracted from the aqueous phase using the MagMAX mirVana Total RNA Isolation Kit (Thermo Fisher, A27828) on a KingFisher Flex Magnetic Particle Processor according to the manufacturer's protocol with 1-2 million cells input. EpiSC: 100-500 ng of total RNA with RIN values of 9.5-10 underwent polyA selection and TruSeq library preparation according to instructions provided by Illumina (TruSeq Stranded mRNA LT Kit), with eight cycles of PCR. Samples were barcoded and run on a NovaSeq 6000 in a PE100 run, using the NovaSeq 6000 S4 Reagent Kit (200 Cycles) (Illumina).  DE: 2 ng total RNA with RNA integrity numbers ranging from 7.7 to 10 underwent amplification using the SMART-Seq v4 Ultra Low Input RNA Kit, with 12 cycles of amplification. Subsequently, 10 ng of amplified cDNA was used to prepare libraries with the KAPA Hyper Prep Kit using eight cycles of PCR. Samples were barcoded and run on a NovaSeq 6000 in a PE100 run, using the NovaSeq 6000 S4 Reagent Kit (200 Cycles) (Illumina).