DNA-seq of multiple specimens
Genomic DNA was extracted using a non-destructive method, combining a home-made lysis buffer with a column-based extraction protocol. A 418 bp fragment of the mitochondrial cytochrome oxidase subunit I (cox1) gene was targeted using the primers BF3 and BR2. To distinguish amplicons from different Malaise traps samples and sites, the primers were tagged with 7 bp tags in the final pooled library. Successful amplified products with different tags on a 96-well plate were pooled and purified using PCR Purification Kit (GeneBetter, Beijing) to construct a single sequencing library. The final three libraries were sequenced on the Illumina NovaSeq 6000 platform with paired-end (PE) 250 bp reads.