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Studying RNA Interactors of Protein Kinase RNA-Activated during the Mammalian Cell Cycle
JoVE Journal
Biyokimya
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JoVE Journal Biyokimya
Studying RNA Interactors of Protein Kinase RNA-Activated during the Mammalian Cell Cycle

Studying RNA Interactors of Protein Kinase RNA-Activated during the Mammalian Cell Cycle

DOI:

10:05 min

March 05, 2019

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Bölümler

  • 00:04Başlık
  • 01:24Preparation of S or M Phase-arrested Cells
  • 02:19Formaldehyde Cross-linking
  • 04:49Immunoprecipitation
  • 07:32Results: Capturing and Analyzing PKR-interacting RNAs During the Cell Cycle Using qRT-PCR
  • 08:45Conclusion

Özet

Otomatik Çeviri

We present experimental approaches for studying RNA-interactors of double-stranded RNA binding protein kinase RNA-activated (PKR) during the mammalian cell cycle using HeLa cells. This method utilizes formaldehyde to crosslink RNA-PKR complexes and immunoprecipitation to enrich PKR-bound RNAs. These RNAs can be further analyzed through high-throughput sequencing or qRT-PCR.

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