1 article published in JoVE
Biotin-based Pulldown Assay to Validate mRNA Targets of Cellular miRNAs Sabyasachi Dash1,2,3, 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.