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The Schlenk tube was capped and the mixture was rapidly stirred at room temperature for 1 min, then was placed into a pre-heated oil bath at the appropriate temperature and stirred for a given time.
The Schlenk tube was capped and the mixture was rapidly stirred at room temperature for 3 min, then was placed into a pre-heated oil bath at 130 °C and stirred for 18 h.
Typically 30 min later, the mixture was rapidly cooled to 0 °C and then the reaction was quenched through the addition of EtOH/HCl (10 wt%).
After addition of the protein solution, the pH of the reaction mixture was rapidly adjusted to 6.4 and maintained constant for 90 min. The reaction was left overnight in the dark at 4°C.
The reaction mixture was rapidly transferred into a cuvette, and absorbance at 420 nm was recorded after every 5 min against the reagent blank prepared simultaneously in the same manner without the analyte.
The mixture was rapidly added to 60 μL of 40 mM AAPH in the well of the microplate and the microplate was immediately placed in the reader, and the fluorescence was recorded at an 485 nm (excitation wavelength) and 535 nm (emission wavelength) every 1 min for 60 min.
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This short length of the MCC ensured that components of the mixture were rapidly separated, i.e. within 70 s.
The reactant mixture expands due to the cracking reaction as gases are formed, and the catalyst/feedstock mixture is rapidly transported up the riser reactor, at speeds approaching 40 m s 1.
The incubated mixtures were rapidly passed through a PBS-washed nitrocellulose filter (diameter 25 mm, pore size 0.45 µm; Sartorius), using a vacuum-assisted filtering apparatus, followed by two washes with 5 ml of PBS.
However, when phenolic substrates (4-tert-butylcatechol or N-acetyl-l-tyrosine) were included in the reaction mixture, procarbazine was rapidly degraded.
Firstly, by adding Triton X-100 and Taq DNA polymerase reaction buffer to the DNA extraction mixture, DNA was rapidly isolated from single diploid cells, single oocytes, early stage embryos and from single hairs.
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