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单分子荧光原位杂交实验中单个大肠埃希菌细胞的转录标记方法

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JoVE Journal Biology

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

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单分子荧光原位杂交实验中单个大肠埃希菌细胞的转录标记方法

DOI:

10.3791/56600-v

•

07:51 min

•

December 21, 2017

•

Rinat Arbel-Goren, Yonatan Shapira, Joel Stavans

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

Automatic Translation

这篇手稿描述了一种方法, 标记个别信使 RNA (mRNA) 转录与荧光标记的 DNA 探针, 用于分子荧光原位杂交 (smFISH) 实验在大肠杆菌. smFISH 是一种可视化方法, 它允许在固定的单个细胞中同时检测、定位和量化单个 mRNA 分子。

Tags

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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