Can I decrease the duration of the hybridisation reaction?
This is not recommended. The hybridisation time should be between 12 and 24 hours for reproducible results. We recommend 16-18 hours, but hybridisation of probes to their targets should be nearly complete after 12 hours. Using more than the recommended amount of probes in order to decrease hybridisation times is not recommended, as it may result in a lower signal and increased background. The use of increased amounts of probes will result in a higher percentage of the unlabeled PCR primer being used for linear amplification of non-ligated probe molecules.
