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Immediately after the addition of the McIlVaine buffer and after incubation for specific times, samples were removed from the reaction mixture for analysis.
CRITICAL STEP Take 12 μl of the overnight RCA reaction mixture for analysis.
Then aliquots were taken from the reaction mixture for analysis [ 14].
Floating cells were retained and added to the adherent cell mixture for analysis.
We vacuum dried 100 µL of the extract and redissolved it in 100 µL of a water/catechol mixture for analysis.
The reaction mixture for analysis consisted of a 25-μl tissue sample, 25 μl of TMB (final concentration, 0.16 mmol/L) dissolved in dimethylsulfoxide, and 200 μl of H2O2 (final concentration, 0.30 mmol/L) dissolved in phosphate buffer (0.08 mol/L, pH 5.4).
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Thin layer chromatography (TLC) is one of the simplest and most widely used methods for separating mixtures for analysis.
For that reason we employ an adaptive two-component mixture model for analysis of the sequence based evidence measure.
We have used around 1% of the ligation mixture for our analysis, a higher amount compared to about 0.02% mentioned for capillary electrophoresis based analysis (15), but using an amount lower than 1% is also possible.
The reaction mixture for ME analysis contained 25 mM malate, 3 mM MgCl2, 80 mM KH2PO4/KOH (pH 7.5), 0.6 mM NADP+ and cell free extract; the reaction was initiated by adding 25 mM malate.
The reaction mixture for HRM analysis consisted of 0.6 μL of each primer (10 μM), 12.0 μL SYBR® Green, 5 ng of genomic DNA and autoclaved nanopure water to a total volume of 20 μL.
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