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Labeling of DNA probes for in situ hybridization is commonly performed by PCR-incorporation of labeled dUTP or dCTP or by an enzyme mixture of DNase I / DNA polymerase I (termed Nick Translation) (Tab. 1, Morrison et al. 2003, Wiegant et al. 2001).   More information...

Check out our enzymatically incorporable Fluorescent, Hapten- , Amine- or CLICK- modified dUTPs or dCTPs as well as corresponding labeling kits!



Figure 1: Chromosome 9 is visualized by fluorescence in situ hybridization (FISH). The ATTO-488-labeled DNA probe was generated from BAC clone BAC268E1 by nick translation with Aminoallyl-dUTP-XX-ATTO-488.

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Table 1: Selected product references. BAC: bacterial artificial chromosome

Product	Enzymatic Method	Template	Reference
Amine Labeling
Aminoallyl-dUTP	Nick Translation	Plasmid DNA	Basu et al. (2014)
Hapten Labeling
Biotin Labeling PCR Core Kit (Biotin-16-dUTP – based)	PCR	Genomic DNA	Garcia et al. (2015)
		PCR fragment	Melnikova et al. (2014)
Digoxigenin-11-dUTP	PCR	chromosomal DNA fragments	Mattern et al. (2015)
		Genomic DNA	Tortajada-Genaro et al. (2016)
Fluorescent Labeling
Aminoallyl-dUTP-ATTO425	Nick Translation	BAC clone	Basinko et al. (2012)
Aminoallyl-dUTP-XX-ATTO488	Nick Translation	BAC clone & genomic DNA	Sanei et al. (2011)
	PCR	chromosomal DNA fragments	Romanenko et al. (2015)
		Whole genome amplified mitochondrial & plasmid DNA	Seok et al. (2015)
		Whole genome amplified BAC clone	Boroviak et al. (2016)
Aminoallyl-dUTP-XX-ATTO532	PCR	single-sorted chromosomal DNA fragments	Potapova et al. (2011)
Aminoallyl-dUTP-ATTO550	PCR	single-sorted chromosomal DNA fragments	Potapova et al. (2011)
Aminoallyl-dUTP-Cy3	Nick Translation	BAC clone and plasmid DNA	Romanenko et al. (2015)
	PCR	chromosomal DNA fragments	Vasconcelos et al. (2016)
		Whole genome amplified BAC clone	Boroviak et al. (2016)
		Whole genome amplified mitochondrial & plasmid DNA	Seok et al. (2015)
Aminoallyl-dUTP-TexasRed	PCR	Whole genome amplified mitochondrial & plasmid DNA	Seok et al. (2015)
		Whole genome amplified BAC clone	Boroviak et al. (2016)
Aminoallyl-dUTP-Cy5	PCR	chromosomal DNA fragments	Romanenko et al. (2015)
		Whole genome amplified mitochondrial & plasmid DNA	Seok et al. (2015)
		Whole genome amplified BAC clone	Boroviak et al. (2016)

Selected References:
Basinko et al. (2012) Clinical and molecular cytogenetic studies in ring chromosome 5: Report of a child with congenital abnormalities. European Journal of Medical Genetics 55:112e.
Basu et al. (2014) Using Amino-Labeled Nucleotide Probes for Simultaneous Single Molecule RNA-DNA FISH. PLOS ONE 9 (9):e107425.
Boroviak et al. (2016) Chromosome Engineering in Zygotes with CRISPR/Cas9. Genesis 54:78
Garcia et al. (2015) Reassignment of Drosophila willistoni genome scaffolds to chromosome II arms. Genes Genomes Genetics 5:2559.
Mattern et al. (2015) Identification of the antiphagocytic trypacidin gene cluster in the human-pathogenic fungus Aspergillus fumigatus. Appl. Microbiol. Biotechnol. 99:10151.
Melnikova et al. (2014) Retrotransposon-Based Molecular Markers for Analysis of Genetic Diversity within the Genus Linum. BioMed Research International 2014:1.
Morrison et al. (2003) Labeling Fluorescence In Situ Hybridization Probes for Genomic Targets. Methods in Molecular Biology 204:21.
Potapova et al. (2011) Loss of centromeric histone H3 (CENH3) from centromeres precedes uniparental chromosome elimination in interspecific barley hybrids. Biotechniques 59 (6):335.
Romanenko et al. (2015) A First Generation Comparative Chromosome Map between Guinea Pig (Cavia porcellus) and Humans. PLOS ONE 10 (5):e0127937.
Sanei et al. (2011) Loss of centromeric histone H3 (CENH3) from centromeres precedes uniparental chromosome elimination in interspecific barley hybrids. PNAS 108 (33):E498.
Seok et al. (2015) Frequent somatic transfer of mitochondrial DNA into the nuclear genome of human cancer cells. Genome Research 25:814.
Tortajada-Genaro et al. (2016) Genotyping of single nucleotide polymorphisms related to attention-deficit hyperactivity disorder. Anal. Bioanal. Chem. 408:2339.
Vasconcelos et al. (2016) Intra- and interchromosomal rearrangements between cowpea [Vigna unguiculata (L.) Walp.] and common bean (Phaseolus vulgaris L.) revealed by BAC-FISH. Genome Research 23:253.
Wiegant et al. (1997) Probe Labeling and Fluorescence In Situ Hybridization. In: Current Protocols in Cytometry 8.3.1, John Wiley & Sons Inc.