title: GSM8016771: PKP2_m1e14_5822_RV; Mus musculus; RNA-Seq
library_construction_protocol: Total RNA was extracted from cardiac RV and LV tissue using the RNeasy Mini or Universal Mini Kit (Qiagen Sciences, 74106 and 73404), cDNA synthesized (Invitrogen SuperScript III First-Strand Synthesis SuperMix for RT) and analyzed by qPCR using Taqman probes to mouse Pkp2 gene, heart failure genes (Nppa, Nppb), and fibrosis genes (Col1a1, Col3a1, and Timp1).  Mouse Gapdh served as an internal housekeeping gene control.  Absolute transgene mRNA copy number was determined by RT-qPCR using a WPRE-specific RNA standard (GenScript) across six orders of magnitude.  DSP protein level in both supernatant and pellet was analyzed to ensure a complete evaluation. From each replicate, 100 ng total RNA was extracted via the polyA-tail-specific protocol according to Illumina Inc.. RNA quality control was performed before library prep using Agilent TapeStation instrument.  The RNA libraries were prepared using a Stranded Total RNA Library Prep with Ribo-Zero Plus kit (Illumina), which also removes ribosomal RNA.  The libraries were sequenced as 2x50 base pair paired-end reads using Illumina NovaSeq 6000 using V1.5 reagent kit on S1 flow cell with an average of 25.66 million reads per each read file (51.32M reads per sample).