GSM7597773: Rat 16; Rattus norvegicus; RNA-Seq
Frozen lung tissue (~35 mg) was homogenized in TRIzol reagent (ThermoFisher Scientific, Waltham, MA). Total RNA was isolated using the Direct-Zol RNA Miniprep Kit (Zymo Research, Irvine, CA) and quantified by a Qubit fluorometer (ThermoFisher Scientific). RNA quality was verified using a Bioanalyzer (Agilent Technologies, Santa Clara, CA). All RNA samples (n =7 – 8 /group) used for sequencing had an RIN ≥ 9.0. RNA samples (1 µg) were processed on an Apollo324 system (Takara Bio USA, Inc. Mountain View, CA). Notably, one sample from the filtered-air exposed group was removed from further processing due to poor quality. Sequencing libraries were prepared using the PrepX RNA-seq Library Kit for Illumina following the manufacturer's protocol. Resulting cDNA libraries were amplified by polymerase chain reaction (PCR) with index primers and subsequently assessed using a Qubit and Bioanalyzer. Pooled libraries were diluted to a final concentration of 1.8 pM + 2% PhiX and then sequenced on the NextSeq 500 platform (Illumina Inc., San Diego, CA).