INTERPRETATION OF HYBRIDISATION RESULTS
The filter is labelled at the top left hand corner
(AI). 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 this filter.
There is one small filter supplied on which 15 plates
have been gridded. As this is the first time that we have provided
this new size of filter we would appreciate if you could let us
know if the use of this filter creates any problems.
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). Library
LL02NP04 contains only 15 plates which occupy panels 1 and 2 only.
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 1-8,2 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:
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. 4-L15). When requesting
this clone it should be preceded by the short library name (e.g.
AI4-L15) and clearly identified as the result of Geneservice filter
screening..
30th Apr. 1997 Version 2.1 (2AI)
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