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Biotin-based Pulldown Assay to Validate mRNA Targets of Cellular miRNAs Video (Video) | JoVE

DOI :

10.3791/57786-v

June 12th, 2018

June 12th, 2018

13,232 Views

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

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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