title: transcriptome of Pimephales promelas: adult amle
design: Briefly, tissues were homogenized in RNA Stat-60 (TelTest) using a handheld rotary homogenizer followed by organic separation with chloroform.  RNA was then subjected to a second round of RNA Stat-60/Chloroform extraction, followed by precipitation in isopropanol overnight at -20C.  The quality of the RNA was assessed using an Agilent Bioanalyzer 2100.  Only samples with RNA integrity numbers (RINs) exceeding 8 were used for sequencing.  Samples were then quantified using a ThermoFisher Scientific Qubit 3.0 fluorimeter. For the PacBio sequencing, an RNA pool was created by adding equal mass of RNA from each of the extracted tissues (brain, liver, gut, testes, heart, gill, head kidney, and trunk kidney) into the pool.