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

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7.1K Views

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10:01 min

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July 30th, 2020

DOI :

10.3791/61520-v

July 30th, 2020

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Sangjin Kim¹^,²^,³^,⁴, Kavya Vaidya¹^,²

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

Podsumowanie

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MRNA Kinetics

Escherichia Coli

Two color SMISH

Single molecule Fluorescence

Transcription Measurement

MRNA Degradation

Time Course Experiments

Cover Slip Preparation

Cell Permeabilization

Rozdziały w tym wideo

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

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