RNA-seq of Leptomonas seymouri grown at 14 degrees C, first replicate
Leptomonas seymouri was cultivated at 23 degrees C in the Brain Heart Infusion medium (Sigma-Aldrich) supplemented with 10 g/ml of hemin (Jena Bioscience), 10% fetal bovine serum, 100 units/ml of penicillin, and 100 g/ml of streptomycin (all three  Sigma-Aldrich). Cells were then seeded at a concentration of 3 x 10^5 cells/ml and cultured for 72 hours at 14 degrees C. Total RNA from three independent biological replicates was isolated from 5  x 10^7 cells using the RNeasy Mini kit (Qiagen) according to the manufacturers instruction. mRNA was prepared in the sequencing center and paired-end strand-specific cDNA libraries were sequenced on Illumina HiSeq platform (Macrogen Inc.).