JoVE Journal

Immunology and Infection

This content is Open Access.

Detection of Bacteria Using Fluorogenic DNAzymes

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).

Capítulos en este video

0:05

Title

2:02

Construction of RFD-EC1 and RFSS1 by Template Mediated Enzymatic Ligation

4:38

Detection using Fluorescence Spectrophotometer

6:28

Detection by Gel Electrophoresis

8:04

Single Cell Detection

9:55

Experimental Validation using E. coli Crude Extracellular Mixtures

12:42

Conclusion

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