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基于生物素的下拉法对细胞 miRNAs mRNA 靶点的验证

JoVE Journal
Genetics

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JoVE Journal Genetics

Biotin-based Pulldown Assay to Validate mRNA Targets of Cellular miRNAs

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基于生物素的下拉法对细胞 miRNAs mRNA 靶点的验证

DOI:

10.3791/57786-v

•

11:00 min

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June 12, 2018

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Sabyasachi Dash^2,3, Muthukumar Balasubramaniam, Chandravanu Dash, Jui Pandhare

¹School of Biotechnology,Kalinga Institute of Industrial Technology University, ²1. Center for AIDS Health Disparities Research Department of Biochemistry and Cancer Biology,Meharry Medical College, ³Kalinga Institute of Industrial Technology University

Chapters

00:04Title
01:34Preparation of Streptavidin Coated Magnetic Beads — I
03:52Preparation of Cell Lysate
05:36Preparation of Streptavidin Magnetic Beads — II
06:34Pull-down of Target mRNA-miRNA Complexes
07:35Total RNA Extracted, cDNA Synthesis, and qPCR
08:57Results: Use of MicroRNAs to Capture the Target mRNA
09:55Conclusion

Summary

Automatic Translation

本报告描述了一种快速可靠的方法来验证细胞 miRNAs 的 mRNA 目标。该方法采用合成生物素化锁定核酸 (低噪声放大) miRNA 模拟捕获靶 mRNA。随后, 采用链亲和素包覆磁性微珠对靶 mRNA 进行 qPCR 聚合酶链反应的定量。

Tags

Biotinylated MiRNA Mimics MRNA Targets Pulldown Assay Locked Nucleic Acids Streptavidin-coated Magnetic Beads MiRNA-mRNA Interactions MiRNA Biology MiRNA Therapeutics

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