I'd really appreciate some assistance with the following question: is it possible to sequentially double digest genomic DNA followed by PCR amplification for the region of interest?
While it makes intuitive sense, my reason for doing this is that I am working with a transgene in a mouse study that is difficult to detect by normal PCR. Thus, my strategy is to increase the PCR efficiency by first setting up a sequential digest with KpnI and ScaI. I'll ethanol purify after each digestion and then will run the PCR. Do you think this will work?
(BTW- I just started the first digest, so any feedback will help me out a lot).