RNA/Zeta Probe Dot Blotting protocol
1) Make up RNA (up to 20ug) dissolved in sterile H2O, TE or 0.5% SDS to 500ul with ice-cold sterile 10mM NaOH, 1mM EDTA and apply it to Zeta Probe membrane, held in a dot-blot apparatus, which has been pre-wetted in 2 x SSC.2) Apply a vacuum across the apparatus to draw the solution through the membrane and wash each dot once with 500ul ice-cold 10mM NaOH, 1mM EDTA.
3) Remove membrane from apparatus and wash for 5 mins in 2 x SSC at room temperature.
4) Allow membrane to air dry.
At this stage the membrane can be stored dry at -20°C until required for hybridisation to the probe of choice or it can be used immediately.
