Hi !!
I m doing luciferase assay for a miRNA-target pair…I m facing a problem…..I hope somebody might help me……
I m co-transfecting miRNA containing plasmid (pSilencer4.1)and pMIR containing the 3’UTR of the target gene….I m finding a decrease in luciferase readings with the increase of miRNA containing plasmid….but the normalization control is also following the same trend…I have tried with 2 different normalization controls-Beta GAl and Renilla…….but I m getting the same results wid both of them…I have also kept the total amount of DNA/PLASMID constant……and the no. of cells in each well is constant……
My boss says she will agree that the miRNA doesnt target the gene we r testing against if I give her constant readings for Luciferase and constant for normalization control….I have repeated this expt enough times but i m not able to figure it out……….plzzzzzzzzzzz help…..