Which is better Biotin Labelling or DIG Labelling kits - (Jun/19/2013 )
I want to do Southern blots.
Have found lot of citations where DIG or Biotin has been used?
Can somebody throw light on pros and con's of using either of the above two methodologies.
In my experience, DIG works better, but the down side is that there are patents owned by Roche, which make reagents etc. much more expensive.
Biotin occurs a lot in natural systems, such as milk. If you block with milk, this will be an issue.
I like DIG too, so long as you realize that the majority of the reagents, other than the anti-DIG antibody and the DIG labeled nucleotides, are easily and relatively cheaply made in the lab, it is quite cost effective and very sensitive.
Really the only reagents you need are the anti-DIG antibody and DIG-dUTP, which you can spike into a PCR reaction master mix. Small amounts go a long way. You have to use a Taq based PCR enzyme because of the U.