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HORST ZITZELSBERGER, BENJAMIN O'BRIEN, and HEINZ-ULRICH WEIER
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Troubleshooting
- Once the probe panel is established and works, make up larger quantities of the probe mix as stock for convenience and consistency,
- If hybridization signals are weak or absent, increase denaturation
times of slides and apply pretreatments like pepsin or proteinase K
digestion; immediately after pretreatment check the slide under the
phase contrast microscope for the presence of any cytoplasm around
the metaphases.
- If swollen chromosomes or disrupted metaphases are present on the
slide, reduce the denaturing time and increase postfLxation after pretreatment procedures. In many cases, aging of slides, e.g. at 37°C for
1 week or at 60°C overnight may also help.
HORST ZITZELSBERGER, BENJAMIN O'BRIEN, and HEINZ-ULRICH WEIER
.. . . fJ
II
•
I"
• ; ...
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~
• ..
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• • •• -
.,.
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11.
.~
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H
1',
Ib
1)
IU
;i
•• •
- --- - ~
- - - - -
'I
1'1
1:0
t:l
.:~ (.
Fig. lb.
Troubleshooting
- Once the probe panel is established and works, make up larger quantities of the probe mix as stock for convenience and consistency,
- If hybridization signals are weak or absent, increase denaturation
times of slides and apply pretreatments like pepsin or proteinase K
digestion; immediately after pretreatment check the slide under the
phase contrast microscope for the presence of any cytoplasm around
the metaphases.
- If swollen chromosomes or disrupted metaphases are present on the
slide, reduce the denaturing time and increase postfLxation after pretreatment procedures. In many cases, aging of slides, e.g. at 37°C for
1 week or at 60°C overnight may also help.