GSM8019693: Kidney, Hp-/-Hpx-/-, 7-months old, Pcc-infected, replicate 3; Mus musculus; RNA-Seq
Total RNA was isolated from mouse organs using tripleXtractor reagent (GRiSP, #GB23.0100), chloroform, isopropanol, and ethanol, according to manufacturer's instructions. Full-length cDNAs and sequencing libraries were generated following the SMART-Seq2 protocol, as previously described (Picelli et al, 2014). Quality control (Agilent Technologies) and "tagmentation" - PCR-mediated adaptor addition and amplification of the adapted libraries was performed following the Nextera library preparation protocol (llumina Tagment DNA Enzyme and Buffer, Illumina #20034211; KAPA HiFi HotStart ReadyMix, Roche #07958935001; Nextera XT Index Kit v2 Set A, Illumina #15052163; Nextera XT Index Kit v2 Set D, Illumina #15052166), essentially as previously described (Baym et al, 2015).  Libraries were sequenced (NextSeq2000 sequencing; Illumina) using 100 SE P2 and sequence information was extracted in FastQ format, using Illumina DRAGEN FASTQ Generation v3.8.4, producing around 25-65 million reads per sample.