title: Illumina MiSeq Sequencing of V3-V4 16S rRNA Amplicons of G. mellonella infected with Heterorhabditis-MA,Nema fraction: day8, bioreplicate2
design: Three healthy G. mellonella larvae were infected and killed with Het-MA nematodes by dousing pieces of Whatman paper with ~ 2,500 infective juveniles in sterile PBS. At 8 days post-death, cadavers were processed by filtering material through a 5um filter, material that did not pass through the filter was resuspended in PBS and DNA extraction was done via methodology recommended by the manufacturer of the PowerSoil kit (MoBio Laboratories, CA, USA). Extracted DNA was quantified with the Quant-iT PicoGreen Assay kit (Life Technologies, NY, USA) and used for V3/V4 16S rRNA locus amplification with custom Illumina barcoded primers based on 341F and 806R sequences.  A paired-end MiSeq run was performed on a 2x250 v2 MiSeq cartridge; the run was 251 cycles (forward) x 12 cycles (index/barcode) and 251 cycles (reverse) using components and methodology according to the manufacturer�s instructions.