The presence of fungi in Armeniacae Semen Amarum, Coicis Semen, Plantaginis Semen, Sesami Semen Nigrum, Ginkgo Semen, Quisqualis Fructus, Chaenomelis Fructus, and Ziziphi Spinosae Semen were analyzed with ITS2(ITS3F_ ITS4R) sequence

This project systematically detected the contaminated fungi of rmeniacae Semen Amarum, Coicis Semen, Plantaginis Semen, Sesami Semen Nigrum, Ginkgo Semen, Quisqualis Fructus, Chaenomelis Fructus, and Ziziphi Spinosae Semen using high-throughput sequencing technologies. Approximately 2g of the EMP samples were transferred into a 15mL sterilized centrifuge tube containing 7 mL of PBS buffer, and the mixture was sonicated for 2 min and shaken with a vortex mixer for 20 s. The preceding extraction steps were repeated, and then the microorganisms were collected from the extraction solution by centrifugation at 13,000 rpm for 10 min. The total DNA was extracted using the soil DNA extraction kitaccording to the manufacturers instructions. ITS2 sequences were amplified with the primer pairs ITS3 and ITS4. The Illumina MiSeq PE300 platform (Illumina, San Diego, CA, USA) was then used to sequence the confirmed PCR products.