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Fluorescence Based Primer Extension Technique to Determine Transcriptional Starting Points and Cleavage Sites of RNases In Vivo
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Fluorescence Based Primer Extension Technique to Determine Transcriptional Starting Points and Cleavage Sites of RNases In Vivo

Fluorescence Based Primer Extension Technique to Determine Transcriptional Starting Points and Cleavage Sites of RNases In Vivo

DOI:
10.3791/52134-v

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10:51 min

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October 31, 2014

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,
1Department of Microbial Genetics, Interfaculty Institute of Microbiology and Infection Medicine Tübingen (IMIT), Faculty of Science,University of Tübingen

Chapters

  • 00:05Title
  • 01:59Primer Extension and Sanger Sequencing
  • 04:26Assembly of Gel Casting Chamber and Gel Casting
  • 06:15Running and Visualizing the Primer Extension Gel
  • 08:41Results: Transcriptional Start Point and RNase Cleavage Site Determination Using Fluorescent Primer Extension
  • 10:20Conclusion

Summary

Automatic Translation

Automatic Translation

We here describe a fluorescence based primer extension method to determine transcriptional starting points from bacterial transcripts and RNA processing in vivo using an automated gel sequencer.

Tags

Keywords: Fluorescence-based Primer ExtensionTranscriptional Starting PointsRNA Processing SitesReverse TranscriptionCDNA FragmentsDenaturing Polyacrylamide Gel ElectrophoresisSanger Sequencing5′-RACENon-radioactiveIn VivoS. AureusE. ColiBacteria

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