Question

Why does Reference strand-mediated conformation analysis (RSCA) needs a fluorophore on its probe? Is it because without the fluorophore probe, the signal (Etbr) would be too weak? Does more mismatches in the heteroduplex causes the ethylBr not bind well and give weak signal and may go undetected in the gel? And how do you order those heteroduplexes in the gel, more mismatches in the heterdoplux->less stable, so less migration and on top of the gel. Less mismatches, more stable heterduplex -> better migration on the bottom of the gel? http://imgur.com/0VP5bjq