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Biochemistry

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An Oligonucleotide-based Tandem RNA Isolation Procedure to Recover Eukaryotic mRNA-Protein Complexes
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An Oligonucleotide-based Tandem RNA Isolation Procedure to Recover Eukaryotic mRNA-Protein Complexes

Article DOI: 10.3791/58223-v • 09:45 min • August 18th, 2018
August 18th, 2018 •
*1, *1, 1
1Dept. of Microbial Sciences, School of Biosciences and Medicine, Faculty of Health and Medical Sciences, University of Surrey
* These authors contributed equally

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résumé
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A tandem RNA isolation procedure (TRIP) for recovery of endogenously formed mRNA-protein complexes is described. Specifically, RNA-protein complexes are crosslinked in vivo, polyadenylated RNAs are isolated from extracts with oligo(dT) beads, and particular mRNAs are captured with modified RNA antisense oligonucleotides. Proteins bound to mRNAs are detected by immunoblot analysis.

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Oligonucleotide-based Tandem RNA Isolation Eukaryotic MRNA-protein Complexes Gene Expression Regulation RNA-binding Proteins Post-transcriptional Gene Regulation Cloning Genetic Manipulation Model Organism Subtype Secondary Structure Analysis Nucleotide Sequence Minimum Free Energy Isolated Base Pairs Interactive RNA Secondary Structure Plot 21 To 24-nucleotide-long Sequences 3-prime Untranslated Regions Guanidine/cytosine Ratio Hairpins Self-annealing 2-prime-methoxy Modified RNA Oligonucleotides Biotin Moiety
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