GSM8861031: 2576-Liver_Mn50; Mus musculus; RNA-Seq
Total RNA from tissue was isolated using the PureLink RNA mini kit using manufacturer's instructions (Invitrogen). Genomic DNA was removed using RapidOut DNA Removal Kit (ThermoFisher). Sequencing libraries were constructed using a 3' Tag-based approach (TagSeq), targeting the 3' end of mRNA fragments. 3' TagSeq was performed by the University of Texas Genomic Sequencing and Analysis Facility (RRID#: SCR_021713) . Libraries were quantified using the Quant-it PicoGreen dsDNA assay (ThermoFisher) and pooled equally for subsequent size selection at 350-550bp on a 2% gel using the Blue Pippin (Sage Science).  The final pools were checked for size and quality with the Bioanalyzer High Sensitivity DNA Kit (Agilent) and their concentrations were measured using the KAPA SYBR Fast qPCR kit (Roche). Samples were then sequenced on the NovaSeq 6000 (Illumina) instrument with single-end, 100-bp reads.