Quality-trimmed bTEFAP sequences of bacterial 16S rRNA from the hind-guts of eight species of carrion beetles (Coleoptera, Silphidae)

DNA from adult hind-guts of 3-6 males and females per species was extracted individually using the SoilMasterTM DNA Extraction Kit (Epicentre, Madison, USA), according to the manufacturer’s instructions (including lysozyme treatment to break up Gram-positive bacterial cells). Identical volumes of the samples for each sex and species, respectively, were pooled for bacterial tag-encoded FLX amplicon pyrosequencing (bTEFAP). DNA was sent to an external service provider (Research & Testing Laboratories, Lubbock, USA) for bTEFAP with 16S rRNA primers Gray28F and Gray519R (Sun et al. 2011) (Table S1). A sequencing library was generated through one-step PCR with 30 cycles, using a mixture of HotStar and HotStar HiFidelity Taq polymerases (Qiagen). Sequencing extended from Gray28F, using a Roche 454 FLX instrument with Titanium reagents and procedures at Research and Testing Laboratory (RTL, Lubbock, TX, USA, http://www.medicalbiofilm.org/). All low quality reads (quality cut-off = 25) and sequences < 200 bp were removed following sequencing, which left 223,470 sequences for analysis (mean ± SD = 13,967 ± 5,946 per sample). Sample abbreviations are as follows: #SampleID Host species NvspFhind Nicrophorus vespillo NvspMhind Nicrophorus vespillo NhumFhind Nicrophorus humator NhumMhind Nicrophorus humator NvesFhind2 Nicrophorus vespilloides NvesMhind2 Nicrophorus vespilloides NorbF_ED Nicrophorus orbicollis NorbM_ED Nicrophorus orbicollis NpusF_ED Nicrophorus pustulatus NpusM_ED Nicrophorus pustulatus NtomF_ED Nicrophorus tomentosus NtomM_ED Nicrophorus tomentosus NameF_ED Necrophila americana NameM_ED Necrophila americana OnovF_ED Oiceoptoma noveboracense OnovM_ED Oiceoptoma noveboracense