Time-course analysis of Drosophila body surface microbiome

Ten female flies of Drosophila melanogaster W1118 each were collected 2, 6 and 10 days post eclosion (DPE). The insects were anesthetized on ice and vortexed with 1 ml PBS buffer for 1 min. The washing buffers were collected and concentrated to 100 microliter by centrifugation. Every four concentrates were pulled together and concentrated to 100 microliter again as one biological repeat. Eight repeats were prepared for each DPE sample. DNA was extracted and amplifed with the 16S V4 primers 515F and 806R. The products were used to generate the amplicon libraries for sequencing with the Illumina Miseq system.