Journal
/
Biologie
/
Probing mRNA Kinetics in Space and Time in Escherichia coli using Two-Color Single-Molecule Fluorescence In Situ Hybridization
Journal JoVE
Biologie
This content is Free Access.
Journal JoVE Biologie
Probing mRNA Kinetics in Space and Time in Escherichia coli using Two-Color Single-Molecule Fluorescence In Situ Hybridization

Probing mRNA Kinetics in Space and Time in Escherichia coli using Two-Color Single-Molecule Fluorescence In Situ Hybridization

DOI:
10.3791/61520-v

•

10:01 min

•

July 30, 2020

•

,
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

Traduction automatique

Traduction automatique

This protocol describes an application of single-molecule fluorescence in situ hybridization (smFISH) to measure the in vivo kinetics of mRNA synthesis and degradation.

Tags

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

Article

Embed

AJOUTER À LA PLAYLIST

Usage Statistics

Vidéos Connexes

check_url/fr/61520?article_type=v

Fluorescence in situ hybridization (FISH) Protocol in Human Sperm
check_url/fr/61520?article_type=v

Analysis of mRNA Nuclear Export Kinetics in Mammalian Cells by Microinjection
check_url/fr/61520?article_type=v

Using Whole Mount in situ Hybridization to Link Molecular and Organismal Biology
check_url/fr/61520?article_type=v

Measuring the Kinetics of mRNA Transcription in Single Living Cells
check_url/fr/61520?article_type=v

A High Throughput in situ Hybridization Method to Characterize mRNA Expression Patterns in the Fetal Mouse Lower Urogenital Tract
check_url/fr/61520?article_type=v

High Throughput Screening of Fungal Endoglucanase Activity in Escherichia coli
check_url/fr/61520?article_type=v

Detection of Viral RNA by Fluorescence in situ Hybridization (FISH)
check_url/fr/61520?article_type=v

Isolation of Ribosome Bound Nascent Polypeptides in vitro to Identify Translational Pause Sites Along mRNA
check_url/fr/61520?article_type=v

Mapping Bacterial Functional Networks and Pathways in Escherichia Coli using Synthetic Genetic Arrays
check_url/fr/61520?article_type=v

Visualization and Analysis of mRNA Molecules Using Fluorescence In Situ Hybridization in Saccharomyces cerevisiae

Read Article