By incorporating 0.1% SDS into the 0.2 M imidazole solution, the contrast
and sensitivity can be even improved.
As an alternative to Zn complexation by citric acid, the gel band can be
incubated ( 2 x 5 min ) in Tris 25 mM, Gly 0.3 M containing 30% ACN, (SDS
is also removed) then wash with 1 ml water for a few seconds. By this way,
there is no change in the pH of the gel during
separation/detection/digestion. We have found very high sequence coverage
following this procedure (Electrophoresis 1999, 20, 732-737)
Lila Castellanos-Serra
Center for Genetic Engineering and Biotechnology
Havana
lila.castellanos@cigb.edu.cu