RNA-Protein Binding Reaction and Separation of Bound RNA
8:35
Reverse Transcription, PCR Amplification, and Protein Binding
9:47
Results: Enrichment of PTB-binding RNAs and Splice Site Switching Assay
10:50
Conclusion
Transcript
The protocol is used for saturation mutagenesis of a protein-binding site. If your protein-binding site is not known, it can be identified using this protocol. The technique has relevance for disease diagnosis and therapy.
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Sequence specificity is critical for gene regulation. Regulatory proteins that recognize specific sequences are important for gene regulation. Defining functional binding sites for such proteins is a challenging biological problem. An iterative approach for identification of a binding site for an RNA-binding protein is described here and is applicable to all RNA-binding proteins.
