Human rna mater panel 2

Total RNA was purified from the tissues according to the manufacturer’s instructions using TRIzol (Life Technologies). For measurements of adult and FCT NPPA and NPPB expression, commercial sources were used as follows: adult heart, ages: 30–39, pooled from 3 male hearts TaKaRa/Clontech “Human RNA Mater Panel II”, #636643, Lot#1208462A); fetal hearts, ages: 21–37wk, pooled from 32 male/female fetal hearts (Clontech “Human Fetal Heart Poly A* RNA”, #636156, Lot#7110214, synthesized with oligo dT20 and SSIII kit). Reverse transcription was achieved using Ready-To-Go You-Prime First-Strand Beads (GE Healthcare, 27-9264-01) following manufacturer’s instructions. Genes were quantified by real-time PCR using SYBR Green primers (Life Technologies) and was carried out on Applied Biosystems Step One Plus. Data analysis was carried out using the log 2-fold change normalized to late-stage week 1 tissue gene expression in Fig. 1b, Extended data Fig. 2a. Data analysis was carried out using the fold change normalized to glyceraldehyde-3-phsophate dehydrogenase (GAPDH) gene expression in Extended data Fig. 2b, 9c. The following primers with confirmed integrity that were used can be found in Supplemental Information Primer List.