title: RNA-seq of Pimephales promelas: adult amle
design: Total RNA was quantitated using a Qubit RNA assay kit and Qubit 2.0 fluorometer (Life Technologies Inc.). 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