Variation of Drosophila ectomicrobiotas after topical challenge with different Metarhizium robertsii strains

The 4 DPE (days post eclosion) female flies of Drosophila melanogaster W1118 were collected, anesthetized on ice and topically infected with the spore suspensions of the wild type and transgenic strains of Metarhizium robertsii. The treated insects were kept at a high moisture condition for 12h. After treatment with each fungal strain, ten flies each were collected, anesthetized on ice and vortexed with 1 ml PBS buffer for 1 min. The washing buffers were concentrated to 100 microliter by centrifugation. Every four concentrates were pulled together and concentrated to 100 microliter again as one biological repeat. Six repeats were prepared for each test sample. DNA was extracted and amplified with the 16S V4 primers 515F and 806R. The products were used to generate the amplicon libraries for sequencing with the Illumina Miseq system.