GSM8752553: N_gru_RNA_Dec_3; Naegleria gruberi; RNA-Seq
DNA from drug-treated cultures were extracted using the NEB Monarch Genomic DNA Purification Kit. Total RNA from these cultures were extracted using the Monarch Total RNA Miniprep Kit. For each library, 100-200 ng of initial gDNA were spiked in with unmethylated lambda genome control and methylated pUC19 plasmid control. These were sheared to 300 bp using a Covaris sonicator, and then used as input for Enzymatic Methyl-Seq kit (New England Biolabs), following manufacturer instructions. We used 200 ng of RNA from treated samples to build mRNA-seq libraries, first enriching for poly-A transcripts with the NEB Magnetic mRNA Isolation Kit S1550S, and then building the libraries with the NEBNext® Ultra™ II Directional RNA Library Prep Kit for Illumina® (E7760L) according to manufacturer's instructions.