The assay mixture was consisted of 1.99 ml 50 mM sodium phosphate buffer (pH 7.0) supplemented with 0.1 μM EDTA, 10 mM guaiacol and 15 mM H2O2 and 100 μl of the enzyme extract in a total volume of 2 ml.
PCR reaction mixture was consisted of 1 μl cDNA, 10 μl pre-mix, 1 μl evergreen fluorescence dye (SolGent, Korea), and 500 nM of each primer (except for 18S reran that 250nM of each primer were used).
The reaction mixture were consist of 0.5 ml enzyme solution and 0.5 ml of 1% colloidal chitosan in 1 ml McIlvaine buffer at the indicated pH.
Heterogeneous asphalt mixtures are consisted of very irregular aggregates, asphalt matrix and a small amount of air voids.
Distinct band with molecular weight of ∼25 kDa was observed in the SDS-PAGE of enterokinase digested mixture, which was consisted with the theoretical molecular weight of CfLec-1.
Therefore, a gas mixture was used consisting of 5% oxygen and 95% nitrogen to decrease the oxygen supply while keeping the aeration rate at 0.013 vvm.
50 μl of reaction mixture was prepared consisting of 50 mM Tris HCl (pH 7.4), 10 mM MnCl2, 5 mM bis-pNPP to which was added 50 μl of the protein sample.
A 6-μl mixture was prepared consisting of 3.5 μl sterile dH2O, 2 μl of restriction enzyme buffer, and 0.5 μl restriction enzymes (HaeIII and MseI).
To calibrate the system, a calibration gas mixture was used consisting of acetaldehyde, acetone, isoprene, benzene, toluene, xylene and α-pinene (covering molecular weights from 32 to 136 amu), each in a concentration of 1 ppmv (parts per million volume, ±5 %) (Linde, Dieren, The Netherlands).
A commercial cellulase mixture was used, consisting of a cellulase derived from Trichoderma reesei (Celluclast 1.5L; Novozymes A/S, Bagsværd, Denmark) (57.8 filter-paper units (FPU /g and 38 IU/g) supplemented with a β-glucosidase preparation (Novozyme 188; Novozymes A/S) (503 β-glucosidase IU/g).
