Table 1.
Forward (F) and Reverse (R) primers for the amplification of 16S rRNA (16S), ITS, rnpB and rpoB fragments for GS-FLX DNA sequencing.
Figure 1.
The number of unique reads for single- and multi-locus sequencing of 16S rRNA.
Comparison of the number of unique 16S rRNA reads generated from samples in which amplicons from four loci were pooled (gray) and those submitted for sequencing singly (black). Bite mark sample 2 contains less than 10 unique reads and was therefore excluded from comparative analyses for all loci.
Figure 2.
The number of unique reads for single- and multi-locus sequencing of ITS.
Comparison of the number of unique ITS reads generated from samples in which amplicons from four loci were pooled (gray) and those submitted for sequencing singly (black). As with 16S rRNA, bite mark sample 2 contains less than 10 unique reads and was therefore excluded from comparative analyses for all loci.
Figure 3.
The number of unique reads for single- and multi-locus sequencing of rnpB.
Comparison of the number of unique rnpB reads generated from samples in which amplicons from four loci were pooled (gray) and those submitted for sequencing singly (black). All rnpB reads were excluded from comparative analyses because bite mark samples 5, 8, 11, 13, 14 contained no reads following quality filtering and bite mark samples 15 and teeth samples 10 and 15 contained less than 10 unique reads.
Figure 4.
The number of unique reads for single- and multi-locus sequencing of rpoB.
Comparison of the number of unique rpoB reads generated from samples in which amplicons from four loci were pooled (gray) and those submitted for sequencing singly (black). Bite mark sample 11 contains less than 10 unique reads and was therefore excluded from comparative analyses for all loci.
Figure 5.
The effect of single- and multi-locus sequencing on unique read numbers.
Comparison of the average number of unique reads generated from samples with varying amounts of DNA, containing either amplicons from one locus or amplicons from four loci.
Figure 6.
Phylogenetic analysis of identical reads shared between bite mark and teeth samples.
All 16S rRNA, ITS and rpoB reads identical between bite mark and teeth samples were compiled into locus-specific files and uploaded into GenBank for standard nucleotide-nucleotide BLAST comparison to determine SLOTUs. The number of shared identical reads in each locus-specific file was 482, 639 and 178, respectively. (Pseudo-Streptococcus pseudopneumoniae; Pneu-Streptococcus pneumoniae; Ther-Streptococcus thermophilus; Vest-Streptococcus vestibularis).
Table 2.
Proportions of unique identical 16S rRNA reads shared between bite mark and teeth samples included in comparative analyses.
Table 3.
Proportions of unique identical ITS reads shared between bite mark and teeth samples included in comparative analyses.
Table 4.
Proportions of unique identical rpoB reads shared between bite mark and teeth samples included in comparative analyses.
Table 5.
The effectiveness of each diagnostic model (i.e. locus) for predicting the probability of obtaining a correct match between a bite mark and the teeth responsible.