Hybridisation
The probe is allowed to bind ("hybridise")
to complementary sequences present in the DNA bands on the blot.
This could be the result:
some bands of the DNA on the blot have been "found" by labelled
DNA from the probe,
which means that they must contain sequences which are also present in the probe
DNA.
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| In the next step, the spots on the blot where label is
present have to be visualised. For this, the Dig label is detected in an enzyme-linked
immunoassay. That's why the Alkaline Phosphatase has been coupled to the
DIG antibody: anti-DIG-AP. With the chromogenic substrate BCIP/NBT the AP can be detected: where the antibody is present, a purple color becomes visible. |