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Genetics

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JoVE Journal Genetics
A Novel Saturation Mutagenesis Approach
 

A Novel Saturation Mutagenesis Approach: Single Step Characterization of Regulatory Protein Binding Sites in RNA Using Phosphorothioates

Article DOI: 10.3791/57816-v • 11:49 min • August 21st, 2018
August 21st, 2018 •
1
1Department of Molecular, Cellular and Developmental Biology, University of Colorado at Boulder

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Proteins that bind specific RNA sequences play critical roles in gene expression. Detailed characterization of these binding sites is crucial for our understanding of gene regulation. Here, a single-step approach for saturation mutagenesis of protein-binding sites in RNA is described. This approach is relevant for all protein-binding sites in RNA.

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Phosphorothioate Approach Saturation Mutagenesis Regulatory Protein Binding Sites RNA Gene Regulation Molecular Biology Single-step Protocol DNA Template Doped Oligonucleotide T7 Primer Non-wild Type Nucleotides Protein-binding Site Doping Site T7 Promoter In Vitro Transcription Phosphorothioate Nucleotide Radiolabeling
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