Place Membranes In A Bottle - Thermo Scientific Shake n Stack 6240 Operating And Maintenance Manual

Place Membranes in
Thermo Scientific
Section 4
Hybridisation
The following procedures indicate how to use the Shake n' Stack Oven for
hybridisation.
1. Place the bottles to be used into the oven during the warm-up period
(approximately 1 hour).
a Bottle
2. Select a piece of support mesh appropriate for the size of the
membrane. The recommended hybridisation mesh is supplied by
Thermo (see parts list in Section 8).
3. Pre-wet the mesh and hybridisation membrane in a suitable tray
containing 2 x SSPE (SSC). See Figure 4-1.
4. Ensuring that the hybridisation membrane exactly overlays the mesh,
roll both up into a tight roll.
If more than one membrane is to be hybridised in a bottle, simply
layer meshes and membranes as required before rolling. It is important
that each membrane is separated from another by a layer of mesh. Up
to five 0.8 x 0.8 in. (20 x 20 mm) membranes can be hybridised in a
single hybridisation bottle.
5. Place 0.34-0.51 fl. oz. (10-15 ml) SSPE (SSC) into a hybridisation
bottle and then insert the roll in such a way that the leading edge
(inside the roll) and the trailing edge are positioned relative to each
other as shown in Figure 4-3.
6. Place the bottle on a flat surface, then slowly unwind the membrane
and mesh around the inside of the bottle by gently rocking and rolling
the bottle along the surface. No air bubbles should be visible between
the membrane and the bottle. If bubbles are present, the membrane
should be removed and re-rolled.
The procedure should then be repeated more gently. Rock the bottle
backwards and forwards to attach the first part of the membrane to the
bottle. Then roll the bottle to unwind the mesh and membrane.
7. Continue until the membrane and mesh are fully unwound. The mesh
ensures the probe will have access to all parts of the hybridisation
membrane including those parts of the membrane which overlap.
Methodology of
Shake 'n Stack 4-1