The DNA library was diluted and the concentration verified using the Qubit assay (Life Technology, Q32850).
The certified concentrations were verified using results from the QX100 droplet digital PCR (ddPCR) system (Bio-Rad).
The concentration was verified using by absorbance at 260 nm using a Nanodrop 2000 with associated software (v. 1.4.1) (Thermo Fisher, Waltham, MA).
Amplicon standards were gel purified (Qiagen), and product length and concentration were verified using a bioAnalyser (model 2100 Agilent Technologies, DNA 1000 LabChip kit).
RNA quality and concentration were verified using an Agilent 2100 bioanalyzer (RNA solutions and RNA 6000 Nano Lab- Chip Kit, Agilent Technologies France, Massy, France).
DNA integrity and concentration were verified using a NanoDrop spectrophotometer (Thermo-Fisher, Pittsburgh, PA) and the Quant-It dsDNA assay (Life Technologies, Carlsbad, CA), which was analyzed on a Qubit fluorometer (Life Technologies).
Blood Alcohol Concentrations (BACs) were verified using infrared spectrometry breath analysis (Intoxilyzer 5000, CMI Inc Ownsboroo, KY, [ 30]).
The satisfactoriness of the numerical methodology used in CFD for obtaining concentration profiles were verified using a classic diffusion problem with its known analytical solution.
Bioethanol concentration was also verified using gas chromatography system (GC17A, Shimadzu, Japan) equipped with flame ionization detector (FID).
In addition, in a separate series of experiments, the bath concentrations of sevoflurane were verified using gas chromatography-mass spectrometry, for sevoflurane concentrations in the perfused gas of 08% 8%, as previously described [ 4].
