Proper controls for RT-PCR - Can we list all the good controls?? (Aug/05/2005 )
Please help me list all the PROPER controls for RT PCR?
When I do my RNA isoloation I also isolate it from a batch that just contains medium, and no cells. This isolate is a good negative control for cross-contamination in the reaction. I always use this as a control when performing RT.
Also, having a household gene, or an RT that has always worked is a good control for the quality/quantity of your RNA isolation. (this last one, I just check when my RT isn't working, then I amplify a piece of a couple 100 bp, which doens't take too long).
Also, if you fear contamination, do all the same steps, but don't add the RT enzyme, and in the end just perform PCR. This way you can check if you have DNA contamination in your RNA isolate.