Kalinga Institute of Industrial Technology University

1 ARTICLES PUBLISHED IN JoVE

Genetics

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

Sabyasachi Dash^1,2,3, Muthukumar Balasubramaniam², Chandravanu Dash², Jui Pandhare²

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.