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INTERPRETATION OF HYBRIDISATION RESULTS
The filter is labelled at the top left hand corner (DROS).
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 is one large filter supplied.
Sheet (i) shows a typical 22 x 22 cm membrane with 36,864
spotted clones. Each filter is divided into 6 panels (indicated by 1,2
......6). The panels are numbered from left to right and eight 384-well
plates are gridded, in duplicate, in each 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 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. Panel
1 contains plates 1-8, panel 2 contains 9-16 and so on. This plate order
has been maintained on all the membranes, e.g.
The interpretation of positive signal(s) can be done by
following the procedure given below:
- Identify the membrane on which the positive signal is located (in
duplicate).
- Use the label position to orientate the filter and identify the
panel in which the positive clonesare located.
- Work out the microtitre co-ordinates of the 4 x 4 array by referring
to the row (A-P) and column (1-24) designations.
- 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:
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Use either table (ii) or table (iii) to identify which
plate number is giving the positive signal.
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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 (e.g. DROS16-L15)
and clearly identified as the result of Geneservice Ltd. filter screening.
Version 1.0 21st Aug. 1997
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