Nick Translation-based DNA Probe Labeling

Labeling of DNA probes for in situ hybridization is most often performed by enzymatic incorporation of labeled dUTP using a mixture of DNase I / DNA polymerase I (Nick Translation) (Tab. 1) (Ribgy et al. (1977), Morrison et al. (2003), Wiegant et al. (2001)).

Figure 1: Metaphase plates of an Arowana fish highlighting the location of 18S rRNA genes after their labeling with ATTO488 NT Labeling Kit through Fluorescence in situ (FISH) experiments. A. DAPI staining. B. 18S rRNA signals C. merged image (courtesy of Prof. Marcelo de Bello Cioffi, Federal University of São Carlos).

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• Rigby et al. (1977) Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. Journal of Molecular Biology 113 (1):237.
• Wiegant et al. (1997) Probe Labeling and Fluorescence In Situ Hybridization. In: Current Protocols in Cytometry 8.3.1, John Wiley & Sons Inc.