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Method for Labeling Transcripts in Individual Escherichia coli Cells for Single-molecule Fluorescence In Situ Hybridization Experiments
 

Method for Labeling Transcripts in Individual Escherichia coli Cells for Single-molecule Fluorescence In Situ Hybridization Experiments

Article DOI: 10.3791/56600-v • 07:51 min • December 21st, 2017
December 21st, 2017 •
1, 1, 1
1Department of Physics of Complex Systems, Weizmann Institute of Science

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This manuscript describes a method for labeling individual messenger RNA (mRNA) transcripts with fluorescently-labeled DNA probes, for use in single-molecule fluorescence in situ hybridization (smFISH) experiments in E. coli. smFISH is a visualization method that allows the simultaneous detection, localization, and quantification of single mRNA molecules in fixed individual cells.

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Keywords: Escherichia Coli Single-molecule Fluorescence In Situ Hybridization (smFISH) Transcripts Cell-to-cell Variability Cell Wall Localization LB Medium Optical Density PBS Fixation Buffer Permeabilization Ethanol Wash Buffer Oligonucleotide Probe Set Hybridization Buffer
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