Genome Sequencing

Firstly, the Y-1 strain was cultured in a liquid ester-producing medium to OD₆₀₀ = 0.7, and the cell genome was extracted by centrifugation. The DNA was extracted using the Omega Fungal DNA Kit D3390-02 according to the manufacturer’s instructions, and NanoDrop was employed to determine the purity and concentration of the samples. The absorbance values at 260 nm and 280 nm were measured. The A₂₆₀/A₂₈₀ ratio was more significant than 1.8, indicating that the nucleic acid was more excellent than 90%, which can be used for subsequent sequencing. Qualified samples were prepared for database construction and quality inspection. The eligible gene library was sequenced, and the final data were subjected to quality control procedures.

DNA degradation and contamination were detected using 1% agarose gel. A nanophotometer was used to determine DNA purity. DNA concentration was determined by TBS-380 fluorometer (Turner BioSystems Inc., Sunnyvale, CA), and the ultrasonic processor lysed the qualified DNA. The inserted fragment was~350 bp in length. Then, end repair, base addition, sequence adapter addition, purification and PCR amplification were performed to complete the preparation of the 350 bp library. The concentration was diluted to 1 ng/μL for the initial quantification with TBS-380 fluorometer. Then, Agilent 2,100 electrophoresis bioanalyzer was used to verify the size of the inserted fragment of the library, and qPCR was performed to measure the effective quantitative concentration of the library. For the identified libraries, sequencing was performed using Illumina HiSeq X Ten, PE150 (Illumina, San Diego, CA, United States).