getting strange data in Luciferase-renilla assayluciferase-renilla
Posted 21 March 2013 - 05:58 PM
I am doing the promega dual luciferase-renilla assay(miRNA testing on UTR)
but my ctrl which for sure was used in other experiments do not go down. Anyway I do not have any desired data about all other samples
renilla reads are really different between different samples but in same cells and I thought it should not differ much between different samples
i got a ratio of Luc per Ren of 1 for my empty vector ctrl but all other samples were higher than 1!
I seeded 70000 cells per well(24well) the next day transfected them and after 24hrs I read the plate
Is there any point that I have missed?any idea?
Posted 22 March 2013 - 10:41 PM
I am not sure what your specific question is. Of course you may have different L/R ratios in different treatments because treatments may altered luc activity. How about the reading of renilla activity alone? Is it pretty consistent across treatments?
Posted 25 March 2013 - 11:17 AM
u know the upregulation was not the expected result as a similar experiment already has done by sb else in our lab.even my positive ctrl that I am sure about its downregulation does not work