title: Ogataea parapolymorpha OppKU80 disruption (A340AA)
design: Genomic DNA of IMD001 was isolated from a stationary overnight culture grown in YPD medium using the QIAGEN Genomic-tip 100/G kit (Qiagen, Hilden, Germany) according to manufacturer’s instructions, and quantified using a Qubit® 2.0 Fluorometer (Thermo Fisher Scientific). 10 µg of genomic DNA was sequenced by Novogene Bioinformatics Technology (Yuen Long, Hong Kong) on a HiSeq 2500 (Illumina, San Diego, CA) with 150 bp paired-end reads using TruSeq PCR-free library preparation (Illumina). In order to verify absence of plasmid pUDP046, the sequencing reads were mapped to reference with pUDP046 and the genome of O. parapolymorpha CBS 11895 (accession GCF_000187245.1)  using the Burrows–Wheeler Alignment tool (BWA) and further processed using SAMtools (Li and Durbin 2010, Li et al. 2009). The absence of sequences from pUDP046 were confirmed by visualizing the generated .bam files in the Integrative Genomics Viewer (IGV) software .