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Metabolisk mærkning af Newly transskriberede RNA til High Resolution genekspressionsprofilering af RNA syntese, videreforarbejdning og Decay i Cell Culture
JoVE Journal
Biology
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JoVE Journal Biology
Metabolic Labeling of Newly Transcribed RNA for High Resolution Gene Expression Profiling of RNA Synthesis, Processing and Decay in Cell Culture
DOI:

11:00 min

August 08, 2013

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Chapters

  • 00:05Title
  • 01:44Metabolic Labeling with 4-thiouridine
  • 03:05Thiol-specific Biotinylation of Newly Transcribed RNA
  • 05:09Separation of Labeled and Unlabeled RNA Using Streptavidin-coated Magnetic Beads
  • 07:00Recovery of Newly Transcribed RNA
  • 08:17Results: Metabolic Labeling of Newly Transcribed RNA
  • 10:25Conclusion

Summary

Automatic Translation

Totalt cellulært RNA giver en dårlig skabelon for at studere kortvarige ændringer i RNA-syntese og forfald samt kinetik RNA forarbejdning. Her beskriver vi metabolisk mærkning af nyligt transkriberede RNA med 4-thiouridine efterfulgt af thiol-specifikke biotinylering og oprensning af nyligt transkriberede RNA gør det muligt at overvinde disse begrænsninger.

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