Remove any air bubbles trapped between the membrane and the filter paper. In all cases, the membrane should not extend beyond the edge of the gasket after the Bio-Dot SF apparatus is assembled. For the best slot blot results, use membrane sheets that have been precut to a 9 x 12 cm size (catalog number 162–0117 for nitrocellulose, 162-0153 for Zeta-Probe membrane). (Touching the membrane to a sheet of filter paper is a simple method for removing excess buffer.) Lay the membrane on the filter paper in the apparatus so it extends over the edges of the filter paper.

Let the excess liquid drain from the membrane. Remove the membrane from the wetting solution. A 10 minute soak is recommended for complete wetting of the membrane to ensure proper drainage of solutions. Introduction The Minifold I manifold is a 96-well filtration/incubation unit for processing dot-blot assays of nucleic acids or proteins. See Sections 9 and 10 for solution preparation. Zeta-Probe membrane is wetted in distilled water. Is wetted in 6x sodium, sodium citrate (SSC) for nucleic acid applications, and in Tris-buffered saline (TBS) for protein binding. Membrane by slowly sliding it at a 45° angle into wetting solution. Always use forceps or wear gloves when handling membranes. Use of Bio-Dot SF filter paper ensures high quality results and eliminates the chance of cross-well contamination.Ĥ. The filter paper is precut to fit inside the sealing gasket. Place the three sheets onto the membrane support. Use the same solution that is used to prewet the membrane (step 4). Moisten three sheets of Bio-Dot SF filter paper (catalog number 162–0161) in wetting Diagram of proper Bio-Dot SF apparatus assembly.ģ.

Slide the plate into the manifold.) Place the sealing gasket on top of the vacuum manifold.įig.1. Place the gasket support plate into position in the vacuum manifold. Clean and dry the Bio-Dot SF apparatus and gasket prior to assembly.Ģ.

Dot blot manifold