INTERPRETATION OF HYBRIDISATION RESULTS

The filter is labelled at the top left hand corner (W). 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 the filter.
There is one small filter supplied.

Sheet (i) shows a 22 x 7.4 cm membrane with 9.408 spotted clones. Filter W consists of two panels (indicated by and ). The panels are numbered from left to right and eight 384-well plates are gridded, in duplicate, in panel and five plates in panel . 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 panel consists of 6144 clones (384 x 8 x 2). Table (ii) shows the order of the plates (in duplicate) within each panel on the filter. Panel contains plates 1-8 and panel contains plates 9-13. This duplicate pattern plate order has been maintained on both panels even when there are less than eight plates gridded, 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. 6-L15). When requesting this clone it should be preceded by the short library name W (e.g. W6L15) and clearly identified as the result of Geneservice filter screening.

Version 1.0 29th March 2001