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Locked Nucleic Acid Flow Cytometry-fluorescence in situ Hybridization (LNA flow-FISH): a Method for Bacterial Small RNA Detection
JoVE Journal
Immunologia e infezioni
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JoVE Journal Immunologia e infezioni
Locked Nucleic Acid Flow Cytometry-fluorescence in situ Hybridization (LNA flow-FISH): a Method for Bacterial Small RNA Detection

Locked Nucleic Acid Flow Cytometry-fluorescence in situ Hybridization (LNA flow-FISH): a Method for Bacterial Small RNA Detection

DOI:
10.3791/3655-v

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09:45 min

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January 10, 2012

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1Center for Bio/Molecular Science and Engineering,Naval Research Laboratory

Capitoli

  • 00:05Titolo
  • 01:25LNA Probes and Experimental Design
  • 02:31Fixation
  • 03:36Diethypyrocarbonate (DEPC) Treatment and Permeabilization
  • 04:53Hybridization and Post-hybridization Washes
  • 05:47Blocking and Staining
  • 07:59Representative Cell Aggregation and LNA flow-FISH Data
  • 09:15Conclusion

Summary

Traduzione automatica

Traduzione automatica

A novel high-throughput method is described that enables the detection and relative quantitation of small RNA and mRNA expression from single bacterial cells using locked nucleic acid probes and flow cytometry-fluorescence in situ hybridization.

Tags

Locked Nucleic AcidFlow CytometryFluorescence In Situ HybridizationFISHNucleic Acid SequencesFlow-FISHLysate-based Nucleic Acid Detection MethodsStatistical AnalysisVariance AnalysisTelomere Length DeterminationCellular IdentificationGene ExpressionViral MultiplicationBacterial Community AnalysisDNA Oligonucleotide ProbesNucleic Acid AnalogsSensitivitySpecificityMelting Temperatures (Tm)Locked Nucleic Acid (LNA) Probes

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