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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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07:51 min

December 21st, 2017

December 21st, 2017

8,084 Views

0:05

Title

0:42

Sample Fixation

2:22

Sample Permeabilization

3:10

Hybridization

4:26

Washing

5:03

Sample Preparation for Imaging

6:12

Results: Representative Results of Two Different Transcripts Labeled by the smFISH Method

7:25

Conclusion

Transcribir

The overall goal of this protocol is to label individual transcripts in E.Coli for fluorescently labeled DNA probes for visualization in single-molecule FiSH experiments. This method can help answer key questions in the systems biology field, such

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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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