We are trying to validate the results of our RNAi screen and we’re wondering what positive and negative controls people generally use to check the efficacy and specificity of the RNAi. We are planning to use the RNAi vector transfected with GFP as a negative control. To check the RNAi effects are specific for the gene we are interested in we were going to construct inserts with smaller pieces of the gene which we hope will confirm the results produced by the RNAi construct containing the whole gene insert.
Does this seem like a valid method? Comments…
sounds like a good plan. the most definitive confirmation of specificity is to get a genetic mutant, ie, order a knock out if one does not exist.
raymond