INTERPRETATION OF HYBRIDISATION RESULTS

The filters are labelled at the top left hand corner (R1 and R2). This label serves as an orientation mark. The enclosed sheets [i, and tables (ii) and (iii)] provide detailed information about the layout of clones on each filter.
There are two small filters supplied, of differing sizes.

Sheet (i) depicts the layout of each filter. Filter R1 has 1 panel (indicated by ) and Filter R2 has 3 panels (indicated by , and ). A full panel (e.g. R2 panel ) has eight 384-well plates gridded, in duplicate. Within each panel, numbers 1-24 correspond to the 24 columns and letters A-P are the 16 rows of the eight 384-well plates in the panel. Each dot represents a clone from a single microtitre plate well. A group of 16 dots corresponds to a 4x4 array of duplicate clones from the 8 microtitre plates (8 x 2). Thus each panel consists of 6144 clones (384 x 8 x 2). Table (ii) shows the order of the 8 plates (in duplicate) within each panel on the filter.
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The 4x4 gridding pattern has been maintained with plates in numerical order. R1 Panel contains plates 1-3, R2 panel contains plates 4-11 and so on. The duplicate spotting plate order has been maintained on all the membranes even where a panel does not contain all 8 plates, e.g.

The interpretation of positive signal(s) can be done by following the procedure given below:

1. Identify the membrane on which the positive signal is located (in duplicate).

2. Use the label position to orientate the filter and identify the panel in which the positive clones
are located.

3. Work out the microtitre co-ordinates of the 4 x 4 array by referring to the row (A-P) and
column (1-24) designations.

4. Within the identified 4 x 4 array determine which plate number contains the positive clone.
Check that any two signals fall in one of the following patterns:

Use either table (ii) or table (iii) to identify which plate number is giving the positive signal.

5. Record the coordinates of putative positives by following the convention
PLATE-ROW and COLUMN (e.g. 16-L15). When requesting this clone it should be preceded by the short library name R (e.g. R16L15) and clearly identified as the result of Geneservice filter screening.

Version 1.0 29th March 2001