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).
Table 1: Overview on available Nick Translation (NT) Kits
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Selected References:
- Chatre et al. (2015) mTRIP: An Imaging Tool to Investigate Mitochondrial DNA Dynamics in Physiology and Disease at the Single-Cell Resolution. In: Weissig V., Edeas M. (eds) Mitochondrial Medicine. Methods in Molecular Biology 1264, Humana Press, New York, NY.
- Himmelbach et al. (2018) Discovery of multi-megabase polymorphic inversions by chromosome conformation capture sequencing in large-genome plant species. The Plant Journal doi:10.1111/tpj.14109.
- Kwiatek et al. (2017) Adaption of the Pivotal-Differential Genome Pattern for the Induction of Intergenomic Chromosome Recombination in Hybrids of Synthetic Amphidiploids within Triticeae Tribe. Frontiers in Plant Science 8 (1300):1.
- Malinovskaya et al. (2018) Copy Number of Human Ribosomal Genes With Aging: Unchanged Mean, but Narrowed Range and Decreased Variance in Elderly Group. Frontiers in Genetics 9 (206):1.
- Matsuyama et al. (2017) Lengthening of high-yield production levels of monoclonal antibody-producing Chinese hamster ovary cell by downregulation of breast cancer 1. Journal of Bioscience and Bioengineering 123 (3):382.
- Mirzaghaderi et al. (2017) Dynamic nucleolar activity in wheat x Aegilops hybrids: evidence of C-genome dominance. Plant Cell Reports 36 (8):1277.
- Morrison et al. (2003) Labeling Fluorescence In Situ Hybridization Probes for Genomic Targets. Methods in Molecular Biology 204:21.
- 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.