title: GSM4474612: Dog 2: Canine primary sublingual epithelial cells, 20 µg/ml D. farinae extract (D2C2); Canis lupus familiaris; RNA-Seq
library_construction_protocol: RNA extraction and DNase treatment of the samples was performed using the RNeasy mini kit (Qiagen) and RNase-Free DNase Set (Qiagen) according to the manufacturer's instructions. Extracted RNA was stored at -80°C. Total RNA concentration was measured using the Quant-iT RiboGreen RNA Assay Kit (Thermofisher Scientific). RNA quality was inspected using an RNA 6000 Nano chip on a BioAnalyser (Agilent Technologies, Santa Clara, CA, USA). For each sample, a sequencing library was constructed using the QuantSeq 3' mRNA-Seq Library Prep Kit FWD for Illumina (Lexogen, Vienna, Austria) starting from 500 ng total RNA and amplified by 14 PCR cycles. Quality and size distribution of the libraries were checked on a BioAnalyser (Agilent Technologies) using the high sensitivity Quant-iT dsDNA Assay Kit (Thermofisher Scientific) and a High Sensitivity DNA chip. Library concentration was determined according to Illumina's Sequencing Library qPCR Quantification Guide on a LightCycler 480 (Roche Life Science, Penzberg, Germany).