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基于寡核苷酸的串联 RNA 分离法恢复真核 mRNA-蛋白复合物
JoVE Journal
Biochemistry
This content is Free Access.
JoVE Journal Biochemistry
An Oligonucleotide-based Tandem RNA Isolation Procedure to Recover Eukaryotic mRNA-Protein Complexes
DOI:

09:45 min

August 18, 2018

Chapters

  • 00:04Title
  • 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

Summary

Automatic Translation

本文介绍了一种用于恢复内在形成的 mRNA 蛋白复合物的串联 RNA 分离过程 (跳闸)。具体来说, rna 蛋白复合物在体内交联, polyadenylated rna 与寡核苷酸 (dT) 珠提取物分离, 特殊基因用修饰 RNA 反义寡核苷酸捕获。通过应用免疫印迹分析, 检测到基因的蛋白质。

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