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En oligonukleotid-baserede Tandem RNA isolering Procedure at inddrive eukaryote mRNA-Protein komplekser
JoVE Journal
Biochemistry
This content is Free Access.
JoVE Journal Biochemistry
An Oligonucleotide-based Tandem RNA Isolation Procedure to Recover Eukaryotic mRNA-Protein Complexes
DOI:

09:45 min

August 18, 2018

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Chapters

  • 00:04Title
  • 00:32Antisense Oligonucleotide (ASO) Design
  • 02:22Protein Extract Preparation
  • 04:08Step 1: Poly(A) RNA Isolation
  • 05:45Step 2: Specific mRNA Capture with 3 -Biotinylated 2 -Methoxy Modified Antisense RNA Oligonucleotides
  • 07:41Results: Capture of Specific mRNA-protein Complexes from Extracts Derived from UV Crosslinked Cells with TRIP
  • 09:00Conclusion

Summary

Automatic Translation

En tandem RNA isolering procedure (tur) inddrivelse af endogent dannes mRNA-protein komplekser er beskrevet. Specifikt, RNA-protein komplekser er crosslinked i vivo, polyadenylated RNA'er er isoleret fra ekstrakter med oligo(dT) perler, og særlig mRNAs er fanget med modificerede RNA antisense oligonukleotider. Proteiner er bundet til mRNAs registreres af immunblot analyse.

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