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Sondering mRNA kinetik i rum og tid i Escherichia coli ved hjælp af to-color single-molecule fluorescens In Situ Hybridization
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Biologie
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Journal JoVE Biologie
Probing mRNA Kinetics in Space and Time in Escherichia coli using Two-Color Single-Molecule Fluorescence In Situ Hybridization

Sondering mRNA kinetik i rum og tid i Escherichia coli ved hjælp af to-color single-molecule fluorescens In Situ Hybridization

DOI:
10.3791/61520-v

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10:01 min

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July 30, 2020

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1Department of Physics,University of Illinois at Urbana-Champaign, 2Center for the Physics of Living Cells,University of Illinois at Urbana-Champaign, 3Carl R. Woese Institute for Genomic Biology,University of Illinois at Urbana-Champaign, 4Center for Biophysics and Quantitative Biology,University of Illinois at Urbana-Champaign

Chapitres

  • 00:04Introduction
  • 00:42Coverslip and Glass Slide Preparation
  • 01:46Time-Course Experiment and Sample Fixation
  • 03:18Cell Membrane Permeabilization
  • 03:57Probe Hybridization
  • 04:31Imaging
  • 05:25Post-Hybridization Wash and Imaging Preparation
  • 06:20Image Analysis
  • 07:36Results: Representative mRNA Kinetic Probing Analysis
  • 09:22Conclusion

Summary

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Traduction automatique

Denne protokol beskriver en anvendelse af enkelt-molekyle fluorescens in situ hybridisering (smFISH) til at måle in vivo kinetik af mRNA syntese og nedbrydning.

Tags

Keywords: MRNA KineticsTwo-color Single-molecule Fluorescence In Situ Hybridization (smFISH)TranscriptionMRNA DegradationEscherichia ColiTime Course ExperimentIPTG InductionONPF RepressionCell FixationCell Permeabilization

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