title: GSM9504605: Ogataea-haglerorum_polyA-mRNAseq_rep4; Ogataea haglerorum; RNA-Seq
library_construction_protocol: One volume of acid phenol (Invitrogen) was added prior to incubation for 60 minutes at 65°C, with vortexing every 10 minutes. Samples were centrifuged for 5 minutes at 16,000 x g at 4°C, and one additional round of acid phenol and one round of chloroform extraction were performed, followed by ethanol precipitation. Samples were treated with DNase I (Sigma-Aldrich) for 15 minutes at room temperature, followed by incubation with 2mM EDTA at 65°C for 10 minutes. Samples were purified using the RNA Clean and Concentrator-25 kit (Zymo Research) according to manufacturer's instructions. Sample quantity was evaluated with a Qubit 3.0 using the RNA Broad Range kit (ThermoFisher) and integrity was evaluated by TapeStation (Agilent) Polyadenylated messenger RNA (polyA mRNA) was selected for high-throughput sequencing with the Universal Plus mRNA-Seq library preparation kit by Tecan per the manufacturer's protocols; triple polyA mRNA-seq replicate libraries were constructed for each strain. All polyA mRNA-seq libraries were Illumina sequenced at the GC3F on a NovaSeq 6000 as SR150 reads.