Southern - How to select probe for such southern analysis? (Jul/03/2002 )
I have done a series of PCR and found that there is slight difference between control and treated group according to bands stained with EB in 2% agarose gel. I was told that such a result is much less sensitive than that of southern. So I blotted such gel to membrane for further southern analysis. Here is the question, if I can use the same PCR product to construct my probe? Yes or No? and How?
Any advice will be appreciated in advance.
So you want to quantitate the PCR product? No need to southernize the DNA. A great method is Pico Green Quanitiation of your PCR products. It will distinguish nicely between unincorporated primers and dNTPs. See Milano et al. Nucleic Acids Research.