Studying reaction kinetics by simultaneous FRET and cross-correlation analysis in a miniaturized continuous flow reactor

摘要: In this study we present a refined optical detection technique for investigation of fast reactions based on confocal fluorescence spectroscopy in miniaturized continuous flow (μCF) reactor. The special setup allows simultaneous observation the reaction basis fluorescent resonance energy transfer (FRET) as an indicator progress. Determination velocity via spatial cross-correlation enables conversion information, i.e. position point, into respective temporal information. To overcome disadvantage conventional reactors high sample consumption, used microfluidic chip allowing economical expenditure solution at low concentration, while silicon/glass is perfectly adaptable to setup. Inside these channels, rapid, diffusion-based mixing under laminar conditions performed crossing three channels by squeezing solutions thin layers. Using μCF device, investigated irreversible cleavage double stranded DNA oligomer enzyme exonuclease III. complementary strands are stained with TAMRA and Cy 5 dye molecules, respectively, undergoing if both annealed. reduction directly corresponds progress reaction.