16S gene of antibiotic treated caddisfly gut microbiome bacterial community
To determine taxonomic and functional differences between bacterial communities present in untreated caddisfly gut microbiomes and gut microbiomes treated with an antibiotic cocktail, we sequenced 16S rRNA. To prepare our samples for sequencing, we added PBS to the gut microbiome sample collected from each caddisfly and filtered this solution through a 13mm 0.22 GPWP01300 filter. These were snap frozen with liquid nitrogen and stored at -80C. DNA was extracted from each filter using the DNEasy Powersoil Pro Kit (Qiagen, Hilden, Germany). We targeted the V4 gene region using the 515f/806r primer. Samples were mutliplexed with a 12 bp tag and sequenced on an Illumina MiSeq instrument set to a PE150 read cycle at Argonne National Laboratory (Lemont, IL, USA).
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AB1