Journal
/
生物化学
/
An Oligonucleotide-based Tandem RNA Isolation Procedure to Recover Eukaryotic mRNA-Protein Complexes
JoVE 杂志
生物化学
This content is Free Access.
JoVE 杂志 生物化学
An Oligonucleotide-based Tandem RNA Isolation Procedure to Recover Eukaryotic mRNA-Protein Complexes

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

10,670 Views

•

09:45 min

•

August 18, 2018

DOI:

10.3791/58223-v

DOI:
10.3791/58223-v

•

09:45 min
August 18, 2018

•

6 Views
, ,
1Dept. of Microbial Sciences, School of Biosciences and Medicine, Faculty of Health and Medical Sciences,University of Surrey

Summary

Automatically generated

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.

Explore More Videos

OligonucleotideTandem RNA IsolationMRNA-protein ComplexesGene Expression RegulationRNA-binding ProteinsPost-transcriptional Gene RegulationEukaryotic MRNASecondary Structure AnalysisBiotinylated OligonucleotidesHEK293 CellsUV Crosslinking

Read Article

Cite this Article

Iadevaia, V., Matia-González, A. M., Gerber, A. P. An Oligonucleotide-based Tandem RNA Isolation Procedure to Recover Eukaryotic mRNA-Protein Complexes. J. Vis. Exp. (138), e58223, doi:10.3791/58223 (2018).

Download .ris file

Copy

Share Video

.

Copy