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Sensitivity analysis indicated that the discrepancies induced by the gas-phase schemes originated either from the presence of heterogeneously-produced water due to its effectiveness as collision partner in the chain terminating reaction H + O2 + M = HO2 + M, or from an overall overprediction of the radical pool in the preignition zone.
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Furthermore, three-body chain terminating reactions involving H2O are enhanced.
The kinetics and specificities of the substrate loading reaction catalyzed by the malonyl/acetyl transferase, the condensation reaction catalyzed by β-ketoacyl synthase and chain-terminating reaction catalyzed by the thioesterase ensure that intermediates do not leak off the enzyme, saturated chains exclusively are elongated and palmitate is released as the major product.
The container was then turned into ice bath to terminate reaction.
When performing Fe VI -US samples taken should be added to a small amount oFe VI -US·L−1 samplestakenulfate to terminate reaction, with the shoulds being addedzed aftor centrifugation.
The terminated reaction was spotted onto a SAM2 Biotin Capture membrane (Promega) and further treated according to the instructions of the manufacturer.
The terminated reactions were processed by conventional SDS-PAGE under denaturing and reducing conditions and transferred to nitrocellulose membranes as described [9].
In all experiments the terminated reactions were analysed on native 20% polyacrylamide gels containing 1× TBE.
Azo reagent NO.1, Azo reagent NO.2 and Azo reagent NO.3 were added to the reaction system successively to terminate reactions.
Separation and quantification of UDP-sugars in terminated reactions was performed by HPAEC analysis using a Dionex Ultimate 3000 system (Thermo Fisher) with detection at 262 nm.
Sulfuric acid (1 N) was added to terminate reactions, and the absorbance at 450 nm was determined by a microtiter plate reader (TECAN, Grödig, Austria).
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