Host species-dependent and seasonally variable microbial communities in the carnivorous pitcher fluids of Sarracenia purpurea and S. psittacina
Each metagenomic DNA sample was subjected to PCR with three sets of primers, each specific for amplification of rRNA sequences from either eubacteria, archaebacteria or eukaryotes. Three replicate PCR reactions were performed for each sample with each primer pair. Replicate reactions were pooled and cleaned by gel extraction of PCR bands with a Qiaquick gel extraction Kit (Qiagen, Valencia, CA). DNA was quantified on an ND-1000 spectrophotometer (NanoDrop, Wilmington, DE) and equimolar quantities of amplicons from each sample were pooled. A single run of Roche-454 Sequencing of 16S eubacterial and 16S archaebacterial amplicon libraries was performed at the Georgia Genomics Facility (University of Georgia, Athens, GA) and 454 sequencing of 18S eukaryotic amplicon libraries was performed at the Interdisciplinary Center for Biotechnology Research, University of Florida, on a 454 Life Sciences Genome Sequencer FLX (Roche) with titanium chemistry.