Influence of cuticular alkanes on the assembly of Drosophila ectomicrobiomes

The study has shown that the P450 gene CYP4G1 contributes to the biosynthesis of cuticular hydrocarbons in Drosophila melanogaster. To determine the gene effect on influencing the assembly of fly cuticular microbiotas, we used the UAS-Cyp4g1 RNAi mutant for investigations by including the W1118 and UAS-mCherry flies as controls. After crossing with the Da-Gal4 flies, the F1 progeny of females was collected three days after eclosion and anesthetized on ice. Ten flies each were vortexed with 1 ml PBS buffer for 1 min, and the washing buffers were concentrated to 100 microliter by centrifugation. Every two concentrates were pulled together and concentrated to 100 microliter again as one biological repeat. Eight 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.