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Table 7 (FISH guide)
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Table 7. Triple color detection scheme for three haptene labeled probes
|
Step |
BIO (blue detection) |
DIG (red detection) |
DNP (green detection) |
|
I. |
Avidin AMCA* |
Mouse antiDIG |
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II. |
Goat antiavidin BIO* |
Sheep antimouse-Cy3 |
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III. |
Avidin AMCA |
Donkey antisheepCy3 |
Rabbit antiDNP |
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IV. |
Mouse antiavidin BIO |
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Goat antirabbit-FITC |
|
V. |
Avidin AMCA |
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Donkey antigoat-DTAF |
* first two steps in BIO detection are optional.
(Note: when using AMCA, DAPI counterstaining of chromosomes is not possible, as these two dyes have similar absorption and emission spectra)