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An Oligonucleotide-based Tandem RNA Isolation Procedure to Recover Eukaryotic mRNA-Protein Complexes
章
- 00:04標題
- 00:32Antisense Oligonucleotide (ASO) Design
- 02:22Protein Extract Preparation
- 04:08Step 1: Poly(A) RNA Isolation
- 05:45Step 2: Specific mRNA Capture with 3 -Biotinylated 2 -Methoxy Modified Antisense RNA Oligonucleotides
- 07:41Results: Capture of Specific mRNA-protein Complexes from Extracts Derived from UV Crosslinked Cells with TRIP
- 09:00Conclusion
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.
