Articles by Sergio D. Aguirre in JoVE

Detection of Bacteria Using Fluorogenic DNAzymes

JoVE 3961 5/28/2012

Sergio D. Aguirre¹, M. Monsur Ali¹, Pushpinder Kanda¹, Yingfu Li^1,2

¹Department of Biochemistry and Biomedical Sciences, McMaster University, ²Department of Chemistry and Chemical Biology, McMaster University

We have recently reported a novel approach for generating fluorogenic DNAzyme probes that can be applied to set up a simple, "mix-and-read" fluorescent assay for bacterial detection. These special DNA probes catalyze the cleavage of a chromophore-modified DNA-RNA chimeric substrate in the presence of crude extracellular mixture (CEM) produced by a specific bacterium, thereby translating bacterial detection into fluorescence signal generation. In this report we will describe key experimental procedures where a specific DNAzyme probe denoted "RFD-EC1" is employed for the detection of the model bacterium, Escherichia coli (E. coli).