ReadiLink™ iFluor® 555 Nick Translation dsDNA Labeling Kit

ReadiLink™ iFluor® 555 Nick Translation dsDNA Labeling Kit is an orange-fluorescent DNA labeling kit designed for enzymatic incorporation of iFluor® 555 into double-stranded DNA probes.

Labeling Principle: Uses DNase I and DNA Polymerase I to incorporate iFluor® 555-dUTP.
Spectral Properties: Excitation/emission maxima of 557/570 nm.
Sample Compatibility: Supports genomic DNA, BAC DNA, PCR products, and plasmid DNA.
Applications: Suitable for preparing fluorescent DNA probes for fluorescence in situ hybridization (FISH).

Nick translation labeling of DNA starts with the creation of defects within the sequence of existing DNA double-helix molecules by cleavage of phosphodiester bonds with DNase along the backbone of one strand. Polymerase then repairs these nicks beginning with the removal of the adjacent nucleotide and the immediate filling back in of those gaps with new nucleotides from the added dNTP pool. As each new nucleotide is added, the polymerase leaves the 3′ OH group open, thus translating the nick toward the 5′ end. As the reaction sequence is repeated, the polymerase enzyme continues to remove existing nucleotides and replace them with new ones at the site of the new nick. The result of these reactions is numerous labeled and unlabeled nucleotides being incorporated as a complementary sequence along the length of each DNA strand, starting at the site of the original nick.

Protocol

Catalog	Size	Price	Quantity
17402	10 Reactions	Price
17403	20 Reactions	Price

Overview

ReadiLink™ iFluor® 555 Nick Translation dsDNA Labelling Kit provides a simple and efficient way to label a double stranded DNA sample with the bright and photostable iFluor® 555 dye. The labelling kit provides all necessary reagents for a complete workflow required for DNA labelling. This method utilizes a combination of DNAse and DNA polymerase to nick one strand of the DNA helix, to which iFluor® 555 dye is conjugated. In addition, the kit allows the user to optimize incorporation and product size by adjusting the ratio of iFluor® 555-dUTP conjugate to dTTP. It is compatible with a wide variety of sample materials, including bacterial artificial chromosome (BAC) DNA, human genomic DNA, purified PCR products, supercoiled and linearized plasmid DNA. The resulted iFluor® 555-labeled DNAs can be used in a variety of molecular biology techniques such as fluorescence in situ hybridization (FISH).

Components

Catalog 17402 (10 Reactions)

Component A: iFluor® 555-dUTP	1 vial (20 µL)
Component B: DNA Polymerase I	1 vial (5 µL)
Component C: DNase I	1 vial (10 µL)
Component D: dNTP mix (dATP, dGTP, dCTP)	1 vial (50 µL)
Component E: dTTP	1 vial (20 µL)
Component F: Nick Translation Buffer	1 vial (100 µL)
Component G: Stop Solution	1 vial (100 µL)

Catalog 17403 (20 Reactions)

Component A: iFluor® 555-dUTP	2 vials (20 µL/vial)
Component B: DNA Polymerase I	2 vials (5 µL/vial)
Component C: DNase I	2 vials (10 µL/vial)
Component D: dNTP mix (dATP, dGTP, dCTP)	2 vials (50 µL/vial)
Component E: dTTP	2 vials (20 µL/vial)
Component F: Nick Translation Buffer	1 vial (100 µL)
Component G: Stop Solution	1 vial (100 µL)

References

View all 6 references: Citation Explorer

Engineering nicking enzymes that preferentially nick 5-methylcytosine-modified DNA.

Authors:

Gutjahr, Alice and Xu, Shuang-yong

Journal:

Nucleic acids research (2014): e77

Pro-apoptotic gene knockdown mediated by nanocomplexed siRNA reduces radiation damage in primary salivary gland cultures.

Authors:

Arany, Szilvia and Xu, Qingfu and Hernady, Eric and Benoit, Danielle S W and Dewhurst, Steve and Ovitt, Catherine E

Journal:

Journal of cellular biochemistry (2012): 1955-65

Identification of bacterial cells by chromosomal painting.

Authors:

Lanoil, B D and Giovannoni, S J

Journal:

Applied and environmental microbiology (1997): 1118-23

An evaluation of a new series of fluorescent dUTPs for fluorescence in situ hybridization.

Authors:

Wiegant, J and Verwoerd, N and Mascheretti, S and Bolk, M and Tanke, H J and Raap, A K

Journal:

The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society (1996): 525-9

Directly labeled DNA probes using fluorescent nucleotides with different length linkers.

Authors:

Zhu, Z and Chao, J and Yu, H and Waggoner, A S

Journal:

Nucleic acids research (1994): 3418-22

Spectral properties

Correction factor (260 nm)	0.23
Correction factor (280 nm)	0.14
Extinction coefficient (cm ^-1 M ^-1)	100000 ¹
Excitation (nm)	557
Emission (nm)	570
Quantum yield	0.64 ¹

Usage and storage

Intended use

Research Use Only (RUO)

Instrument settings

Other instruments

Thermal Cycler

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Page updated on November 7, 2026