Bio-Rad Bio-Dot® and Bio-Dot SF Microfiltration Apparatus - Manual (Page 14)

Note: A method for applying gentle vacuum to the apparatus is to adjust the flow valve to
setting 3. Use a finger to cover the valve port exposed to air. The amount of vacuum reaching
the manifold will be regulated by the pressure of your finger on the valve.
6. Rinse all wells to wash any sample on the side of the wells through. Rinse with 500 µl TE.
Apply vacuum (flow valve setting 1, Figure 3) until the sample wells are dry.
7. Disassemble the Bio-Dot SF apparatus. Remove the blotted membrane.
8. Remove glyoxal adducts by pouring 20 mM Tris-HCl, pH 8.0, 1 mM EDTA heated to 95°C
onto the membrane and agitating at room temperature until the solution cools. Alternatively,
place the membrane in a vacuum oven at 80 °C for 1 hour for Zeta-Probe membrane, 2
hours for nitrocellulose, and omit step 8.
9. Nitrocellulose membranes must be baked under vacuum for 2 hours at 80°C before
hybridization. If hybridization is not to be undertaken within 2 days, bake the Zeta-Probe
membrane under vacuum for 30 minutes at 80°C. The Zeta-Probe membrane and
nitrocellulose membranes can be stored dry between two pieces of filter paper in plastic bags
at 23–25°C.
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