GSM1960358: Naked Mole Rat Adult Liver 1; Heterocephalus glaber; RNA-Seq
Harvested livers were preserved in RNAlater (ThermoFisher Scientific) Total RNA quality was checked on an Agilent 2100 Bioanalyzer; only samples with a RNA Integrity Number greater than 8.5 were used for subsequent analysis. Total RNA was treated with DNaseI, column purified using the miRNeasy Mini Kit (Qiagen), and depleted of ribosomal RNA with Ribo-Zero Magnetic Gold Kit (Epicentre), followed by ethanol precipitation. Depleted RNA was converted to cDNA using SuperScript III First-Strand Synthesis Kit (Invitrogen) with 80 ng random hexamers and 50 μM oligo dT and subsequently ethanol precipitated. Single-stranded cDNA was converted to dsDNA by DNA polymerase I while incorporating dU/VTPs (10 mM). Samples were fragmented in 1 × TE to 200–300 bp using Covaris. After fragmentation, samples were purified using the MinElute PCR purification kit (Qiagen). Fragmented samples underwent standard end-repair, dA-tailing and adapter ligation using Illumina TruSeq adapters for multiplexing. Adapter-ligated cDNA was treated with uracil-DNA glycosylase followed by enrichment PCR using Q5 polymerase (New England Biolabs) for 18 cycles. Libraries were size selected for 150–600 bp on a 2% low-melt ultra low-range agarose gel stained with SYBR Gold (Invitrogen) to eliminate adaptor dimers.
GEO Accession
GSM1960358