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At studere RNA Interactors af Protein Kinase RNA-aktiveret under pattedyr cellecyklus
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Chapters
- 00:04Title
- 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 præsenterer eksperimentelle metoder til at studere RNA-interactors af dobbelt-strenget RNA bindende protein kinase RNA-aktiveret (PKR) i løbet af pattedyr celle cyklus ved hjælp af HeLa celler. Denne metode udnytter formaldehyd bitmapgenkendelse RNA-PKR komplekser og immunoprecipitation til at berige PKR-bundet RNA'er. Disse RNA'er kan analyseres yderligere gennem høj overførselshastighed 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
