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During kit validation, a protocol utilizing a 4-hour labeling step and addition of Stop Buffer gave us the best results in terms of dye incorporation, DLR scores, and signal-to-noise ratios. However, omitting the addition of Stop Buffer often does not impede the results significantly and adequate DLR scores can still be obtained. Especially if the DNA is purified immediately afterwards, the stop buffer step and/or heat inactivation may be omitted.