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Chapitres
- 00:04Titre
- 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
Vi presenterar experimentella metoder för att studera RNA-interactmedlemmar dubbelsträngat RNA-bindande protein Kinas RNA-aktiverat (PKR) under mammalian cell cykel använder HeLa cells. Denna metod använder tredjeparts formaldehyd crosslink RNA-PKR komplex och immunoprecipitation att berika PKR-bundna RNAs. Dessa RNAs kan analyseras vidare genom hög genomströmning sekvensering eller qRT-PCR.
Tags
RNA InteractorsProtein Kinase RNA-ActivatedMammalian Cell CycleCrosslinkingFormaldehydeImmunoprecipitationNeurodegenerative DiseaseParkinson’s DiseaseAlzheimer’s DiseaseDouble-stranded RNARNA-binding ProteinsPost-transcriptional RegulationGene ExpressionCell Cycle ArrestHeLa CellsThymidineNocodazole
