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Biological oxygen demand (BOD), chemical oxygen demand (COD), and dissolved organic carbon (DOC) were analysed using the respective standard methods (Eaton et al. 2005).
The results were automatically calculated using the respective standard curves by the LightCycler analysis software version 3.5.
The concentration of each gene was calculated with the aid of the LightCycler software using the respective standard curve as reference.
However, converting back Cq values to molecules using the respective standard curves always reported the number of molecules determined optically at 260 nm indicating that standard curves have the capacity to compensate for primer mismatches.
The integrated areas under the PRO and HYP peaks were divided by the integrated area of the IS and their relative concentrations were obtained using the respective standard curves.
For the WPa-group, the sonic toothbrush (Waterpik® Sensonic SR-3000E) wadjustedted onto the device using the respective standard brush head with a load of the brush head onto the interproximal region of < 0.9 N as measured for sonic toothbrushes (total load 70±± 5 g) [ 2, 16].
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The feasible ranges of the quantified intracellular metabolites were calculated using the respective experimental mean and standard deviation.
AS and ASL, and their para- (ASpOH, ASLpOH) and ortho-hydroxy-metabolites (ASoOH, ASLoOH), were determined by LC-MS-MS analysis using the respective deuterium labeled analogues as internal standards, essentially as described [ 38].
mRNA copy numbers were calculated using standard curves constructed using the respective PCR products eluted from agarose gels.
Both strands of all PCR products were sequenced, using the respective primer pair used for amplification and standard vector primers M13F and M13R for the cloned products, employing the BigDye™ Terminator v1.1 Cycle Sequencing Kit (Applied Biosystems) on an ABI Prism™ 3100-Avant-Genetic 3100-Avant-Genetic 3100-Avant-Genetic
The concentration of CXCL13 in the sera was interpolated from a standard curve, which was generated using the respective recombinant protein.
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