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

DOI:
10.3791/56600-v

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07:51 min

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December 21, 2017

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, ,
1Department of Physics of Complex Systems,Weizmann Institute of Science

Chapters

  • 00:05Title
  • 00:42Sample Fixation
  • 02:22Sample Permeabilization
  • 03:10Hybridization
  • 04:26Washing
  • 05:03Sample Preparation for Imaging
  • 06:12Results: Representative Results of Two Different Transcripts Labeled by the smFISH Method
  • 07:25Conclusion

Summary

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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.

Tags

Keywords: Escherichia ColiSingle-molecule Fluorescence In Situ Hybridization (smFISH)TranscriptsCell-to-cell VariabilityCell Wall LocalizationLB MediumOptical DensityPBSFixation BufferPermeabilizationEthanolWash BufferOligonucleotide Probe SetHybridization Buffer

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