Illumina sequencing of Acetobacter Tropicalis: Drosophila melanogaster
Five whole-body Drosophila park25 mutants were homogenized and plated on modified MRS medium. Copper and tan colonies were streaked for isolation and their taxonomic assignment as Acetobacter species was confirmed by Sanger sequencing. DNA was extracted using the Qiagen DNeasy Powerlyzer Microbial Kit, fragmented to 500bp (NEBNext dsDNA Fragmentase, M0348L), end repaired (NEBNext Ultra II End Repair/dA-Tailing module, E7546L), ligated to Illumina sequencing adapters (NEBNext Ultra II Ligation Module, E7595L), and size selected to ~500bp. Ligated target sequences were enriched with the KAPA Library Amplification kit (KK2621), size selected to 500bp, and normalized to 5ng/uL using Qubit measurements.