GSM3036896: PK6-2: Ad-Cre PNPT1 fl/fl (PKO MEF) Clone 6 Replicate 2; Mus musculus; RNA-Seq
TM6, rho0 MEFs, and PKO clones were grown in biological triplicates to 70-80% confluence and purified using TriZol Reagent (Life Technologies) or RNeasy Mini Kit (Qiagen). All samples used showed a A260/280 ratio > 2.00 (Nanodrop; Thermo Scientific). Strand-specific ribosomal RNA (rRNA) depleted RNA-Seq libraries were prepared from 1 µg of total RNA using the KAPA Stranded RNA-Seq Kit with Ribo-Erase (Kapa Biosystems, Roche). Briefly, rRNA was depleted from total RNA samples, the remaining RNA was heat fragmented, and strand-specific cDNA was synthesized using a first strand random priming and second strand dUTP incorporation approach. Fragments were then A-tailed, adapters were ligated, and libraries were amplified using high-fidelity PCR. All libraries were prepared in technical duplicates per sample (n = 12 samples, 24 libraries total), and resulting raw sequencing reads merged for downstream alignment and analysis. Libraries were paired-end sequenced at 2x125bp on an Illumina HiSeq.
GEO Accession
GSM3036896