Apogee 21054010 Buku Petunjuk - Halaman 5

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2.0 INSTRUCTIONS FOR USE

Refer to Figure 1 to identify the features and components in these instructions.

The capillary blot technique originally developed by Southern (1) has been modified and extended in a

number of ways since its original publication. Protocols with buffer compositions for DNA and RNA blots

can be found in a number of journal articles and laboratory manuals (2 thru 5).

All protocols require one or more soaking steps before actual blotting begins. During these steps, the

agarose gels are fragile and easily damaged; whenever possible, handle the gel using the UVT tray on

which it was cast.

To help prevent the gel from slipping off the UVT tray during handling, place the gel restrainer over the

UVT tray. Orient the gel restrainer so that the bent tabs block the open ends of the UVT tray (Figure 2).

All soaking steps can be carried out in the buffer tray. Transfer the gel, in the UVT tray, into the buffer

tray and cover the gel completely with the appropriate soaking solution. Rock the buffer tray and gel

gently and regularly during soaking steps. Remove the gel and thoroughly rinse the buffer tray with

deionized water before adding each new solution.

Figure 2: Gel restrainer in position over UVT tray

1. When all soaking steps are complete, remove the gel, in the UVT tray or gel plate, to a clean

work surface. Use the gel restrainer as needed while handling the gel in the UVT tray.

2. Wet a wicking sheet in the solution to be used for capillary transfer.

3. Place the wicking sheet on the gel. If using a UVT tray, align the edges of the wicking sheet

against the side rails of the tray with the overlap of the wicking sheet equal at the top and

bottom of the gel. The wicking sheet should run 90 degrees to the side rails of the UVT tray so

that the overlap does not run up the side rails.

Note: Do not trap air pockets between the gel and wicking sheet. Remove trapped air by lifting and

replacing the wicking sheet or by gently finger. Small bubbles in the wells will not affect transfer.

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