Fig 1.
Schematic overview of the quantitative sequencing method and normal illumina sequencing workflows.
Fig 2.
Plot of expected copy number versus copy number determined experimentally by qSeq.
Genomic DNA of S. tokodaii was used for the test. Error bars indicate standard deviation from triplicate measurements. Understimation of DNA was obseved when template DNA is more than 1.0 × 105 copies.
Table 1.
Quantification of 16S rRNA gene in the mock microbial community.
Fig 3.
Analysis of mock microbial communities.
(A) Mock microbial community of 6 micriboal species. Relative abundances were detemined by qSeq and standard illumina sequencing library. Standard deviation was calcurated from triplicate measurements. (B) Mock microbial community of 16S rRNA genes of HE and MJ at different ratios. Plot shows MJ to ME measured ratios versus expected ratio. (C) 16S rRNA genes of SA and BS at different ratios. Plot shows SA to BS measured ratios versus expected ratio. Error bars indicate standard deviation from triplicate measurements. ST, S. tokodaii; MJ, M. jannaschii; BS, B. subtilis; SA, S. avermitiis; PD, P. denitrificans; HE, H. elongata.
Fig 4.
Analysis of environmental samples.
(A) 16S rRNA genes were quantified by dPCR and qSeq using DNA extracted from the environmental samples. Error bars indicate standard deviation from triplicate measurements. (B) Ratio of archaea to bacteria determined by dPCR, qSeq and the standard Illumina sequencing library. Error bars indicate standard deviation from triplicate measurements. As for biofilm and hot spring sample, the ratio was calculated from single sample.
Table 2.
Quantification of the copy number 16S rRNA gene of selected classes.
Table 3.
Sequences of oligonucleotide primers used in this study.