Total RNA was isolated from each population using the Agencourt RNAdvance Cell Kit (Beckman Coulter). Samples were processed starting with 2ng total RNA. cDNA libraries were generated using the Single Primer Isothermal Amplification (SPIA) RNA-Seq protocol. Libraries were quantitated by Qubit and qPCR and one equimolar pool was made based upon qPCR results. Following denaturation 2.0pM of library pool was clustered on a high output NextSeq run (75bp SR sequencing using Illumina NextSeq550 Hihg Output mode).