A series of two fold dilutions of each extract in the final concentration of 512, 256, 128, 64 and 32 μg/ml were prepared in MH agar.
In a further 27 patients, stones were successfully extracted in the second session of treatment (final success rate: 92.7%).
Cellular proteins were extracted in a final volume of 250 μL in 100 mM Tris pH 7.8, 200 mM NaCl, 1 mM EDTA, 2% (v/v) β-Mercaptoethanol and 0.2% (v/v) Triton X-100 by sonication.
Chlorophylls and carotenoids were extracted in acetone (80% final concentration buffered with Na2CO3) and measured by fitting of the absorption spectrum of acetone extracts [ 79].
The 1000-bp upsequencesquences of PP2C genes in subfamily A were extracted from the final databases from Arabidopsis and rice.
The reaction mixture contained mouse liver homogenate 0.1 ml (25%, w/v) in Tris HCl buffer (20 mM, pH 7.0), KCl (150 mM), FeSO4 · 6H2O (0.8 mM), ascorbic acid (0.3 mM) and various concentrations of the extract in a final volume of 0.5 ml and was incubated for 1 h at 37°C.
To perform the activation of cryptic trehalase, 50 μL of the activation cocktail containing 2 mM ATP, 20 mM MgCl2, 50 μM cAMP, 50 mM NaF and 5 mM theophylline prepared in 50 mM phosphate buffer, pH 7.5, was added to a suitable sample of the crude extract in a final volume of 100 μL.
A set of SNPs on an S-gene was defined as "gene-based eQTL" (g-eQTL) if the number of extracted genetic factors in the final model was greater or equal to 1, and the variation in the S-gene and T-gene accounted for by the factor(s) was non-zero.
Briefly, the reaction mixture contained 1× TaqMan Universal PCR Master Mix Applied Biosystemss), 300 400 nM of each primer, 200 nM of each probe, and 5μl of DNA extract in a final volume of 25 μl.
Typical enzyme assay conditions were 40 mM L-aspartate, 0.4 mM carbamoyl phosphate containing [C]carbamoyl phosphate (0.1 μCi μmole -1 ), Tris buffer (0.1 M, pH 8.0), and cell-free extract in a final volume of 200 μl.
