16s amplicon sequencing of DNA extracted from Anopheles stephensi mosquitoes
Library was prepared by two-stage PCR, targeting the V4 and V5 variable regions of the microbial small subunit ribosomal RNA gene. The pooled library was purified using an AMPure XP cleanup protocol (0.6X, vol/vol; Agencourt, Beckmann-Coulter) to remove fragments smaller than 300 bp. The pooled libraries, with a 20% phiX spike-in, were loaded onto an Illumina MiniSeq mid-output flow cell (2x150 paired-end reads).