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

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

English

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13,425 Views

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11:00 min

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June 12, 2018

DOI:

10.3791/57786-v

DOI:
10.3791/57786-v

•

11:00 min
June 12, 2018

•

7 Views
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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

Summary

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This report describes a fast and reliable method for validating mRNA targets of cellular miRNAs. The method uses synthetic biotinylated Locked Nucleic Acid (LNA)-based miRNA mimics to capture target mRNA. Subsequently, streptavidin-coated magnetic beads are employed to pulldown the target mRNA for quantification by qPCR polymerase chain reaction.

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Biotinylated MiRNA MimicsMRNA TargetsPulldown AssayLocked Nucleic AcidsStreptavidin-coated Magnetic BeadsMiRNA-mRNA InteractionsMiRNA BiologyMiRNA Therapeutics

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Cite this Article

Dash, S., Balasubramaniam, M., Dash, C., Pandhare, J. Biotin-based Pulldown Assay to Validate mRNA Targets of Cellular miRNAs. J. Vis. Exp. (136), e57786, doi:10.3791/57786 (2018).

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