ITS sequencing of soil samples with biochar and microplastics application
The primers of ITS1F/ITS2R were applied to amplify the fungal ITS gene for the fungal community. The PCR products were purified with AMPure XT beads (Beckman Coulter Genomics, Danvers, MA, USA) and quantified by Qubit (Invitrogen, USA). The samples were sequenced on an Illumina NovaSeq platform at LC-Biological Technology Co. Ltd to generate paired-end reads assigned to samples based on their unique barcode. Paired-end reads were merged using FLASH. The raw reads were filtered under specific conditions for high-quality clean tags according to the fqtrim (v0.94). Chimeric sequences were filtered using Vsearch software (v2.3.4). Following dereplication, quality control and identification of amplicon sequence variants were conducted using DADA2.