RAY@leicester.ac.uk (02/01/90)
>I have a small technical problem, perhaps one of you can give me a simple >solution. > >When I electrophorese my pcr products, I am looking for a band around >200 bases long (pretty small). The problem is that the band usually comes >out right at the dye front, which makes UV visualization a slight problem. > >Is there another dye I can use that will run a bit faster or slower than >bromphenol blue? I could try just using glycerol and edta in my loading >buffer, leaving out the dye. > >All suggestions welcome... > >Daniel G. Sinclair >Rsch Tech II >University of NC, Chapel Hill > There certainly is an alternative to bromophenol blue. Orange G (available form Sigma cat no. O1625) is much better. It runs faster than BPB and tends to quench UV visualisation much less. The amount to use is unimportant. Just add enough so that the loading dye looks orange. For the perfectionists that's about 0.1% w/v in a 10x loading dye. Raymond Dalgleish, Department of Genetics, University of Leicester.