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

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10:01 min

July 30th, 2020

July 30th, 2020

6,796 Views

0:04

Introduction

0:42

Coverslip and Glass Slide Preparation

1:46

Time-Course Experiment and Sample Fixation

3:18

Cell Membrane Permeabilization

3:57

Probe Hybridization

4:31

Imaging

5:25

Post-Hybridization Wash and Imaging Preparation

6:20

Image Analysis

7:36

Results: Representative mRNA Kinetic Probing Analysis

9:22

Conclusion

Transcript

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

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