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Detection of Bacteria Using Fluorogenic DNAzymes
JoVE Journal
Immunology and Infection
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JoVE Journal Immunology and Infection
Detection of Bacteria Using Fluorogenic DNAzymes
DOI:

13:20 min

May 28, 2012

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Chapters

  • 00:05Title
  • 02:02Construction of RFD-EC1 and RFSS1 by Template Mediated Enzymatic Ligation
  • 04:38Detection using Fluorescence Spectrophotometer
  • 06:28Detection by Gel Electrophoresis
  • 08:04Single Cell Detection
  • 09:55Experimental Validation using E. coli Crude Extracellular Mixtures
  • 12:42Conclusion

Summary

Automatic Translation

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).

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