V4 16S  Periplaneta americana microbiome: foregut
Cockroach gut samples were placed in 800uL 1X PBS upon dissection and disrupted with a sterile pestle. Microbial DNA was extracted (Omega Biotek Bacterial DNA kit) from 200ul aliquots of suspended lumen material (sans gut tissue) and quantitated, and 4ng of DNA was used as input for 2-step PCR barcoding targeting the V4 region of the 16S rRNA gene. Amplification was verified via gel electrophoresis, and PCR products were cleaned (Omega Biotek Cycle Pure kit) and quantitated before pooling together to prepare an equimolar sequencing library.