DNA-Seq of Talaromyces marneffei: internal transcribed spacer region
DNA extraction was achieved by firstly disrupting fungal cells with glass beads using TissueLyser II and then the released fungal DNA was purified using the QIAquick PCR Purification Kit according to the manufacturers protocol. One microlitre of the extracted DNA was used for PCR-amplification. PCR amplification of the ITS of the two isolates were performed according to a published protocol, using ITS1 (5-TCCGTAGGTGAACCTGCGG-3) and ITS4 (5-TCCTCCGCTTATTGATATGC-3) as the PCR primers. DNA sequencing was performed by next-generation sequencing using the Oxford Nanopore Technologies MinION device. The sequences of the PCR products were compared with sequences of closely related species in the DDBJ/ENA/GenBank databases (core nucleotide database) by pairwise alignment using nucleotide BLAST.