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siRNA Transfection and EMSA Analyses on Freshly Isolated Human Villous Cytotrophoblasts
Capítulos
- 00:05Título
- 01:00Evaluation of the Purity of Isolated Primary Villous Cytotrophoblasts
- 03:13siRNA Transfection of Human Primary Villous Cytotrophoblasts
- 04:55Preparation of Nuclear Extracts from Cultured Cells
- 05:52Radiolabeling of Single Stranded Oligonucleotide
- 06:50DNA Binding Reaction
- 08:07Results: Syncytin-2 Knowckdown and Visualization of Syncytin-2 DNA-protein Complexes
- 09:14Conclusion
This protocol describes a method for efficiently transfecting siRNA in freshly isolated human villous cytotrophoblasts using microporation and identifying DNA-protein complexes in these cells. Transfected cells can be monitored by Western blot and EMSA analyses during the 4-day culture time.
