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利用两色单分子荧光在原位杂交中
Escherichia coli
探索大肠杆菌中空间和时间中的 mRNA 动力学
JoVE Journal
Biology
This content is Free Access.
JoVE Journal
Biology
Probing mRNA Kinetics in Space and Time in
Escherichia coli
using Two-Color Single-Molecule Fluorescence In Situ Hybridization
Please note that all translations are automatically generated.
Click here for the English version.
利用两色单分子荧光在原位杂交中
Escherichia coli
探索大肠杆菌中空间和时间中的 mRNA 动力学
DOI:
10.3791/61520-v
•
10:01 min
•
July 30, 2020
•
Sangjin Kim
2,3,4
,
Kavya Vaidya
2
1
Department of Physics
,
University of Illinois at Urbana-Champaign
,
2
Center for the Physics of Living Cells
,
University of Illinois at Urbana-Champaign
,
3
Carl R. Woese Institute for Genomic Biology
,
University of Illinois at Urbana-Champaign
,
4
Center for Biophysics and Quantitative Biology
,
University of Illinois at Urbana-Champaign
Chapters
00:04
Introduction
00:42
Coverslip and Glass Slide Preparation
01:46
Time-Course Experiment and Sample Fixation
03:18
Cell Membrane Permeabilization
03:57
Probe Hybridization
04:31
Imaging
05:25
Post-Hybridization Wash and Imaging Preparation
06:20
Image Analysis
07:36
Results: Representative mRNA Kinetic Probing Analysis
09:22
Conclusion
Summary
Automatic Translation
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Automatic Translation
该协议描述了单分子荧光原位杂交(smFISH)用于测量mRNA合成和降解的体内动力学的应用。
Tags
MRNA Kinetics
Two-color Single-molecule Fluorescence In Situ Hybridization (smFISH)
Transcription
MRNA Degradation
Escherichia Coli
Time Course Experiment
IPTG Induction
ONPF Repression
Cell Fixation
Cell Permeabilization
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