Here, we used RNA-seq to quantify expression conservation across many Ascomycete yeasts. Overall design: We used strand-specific RNA-seq (dUTP) to quantify mRNA expression in each species. There was one replicate per species, and all species were grown in BMW media to log phase. With the exception of N. castellii (25C), all others were grown at 30C. Expression conservation was calculated using the RPKM for each species using orthologous genes.