ReadiLink™ Cy5 Nick Translation dsDNA Labeling Kit

Name: ReadiLink™ Cy5 Nick Translation dsDNA Labeling Kit
Brand: AAT Bioquest
SKU: 17408
Availability: InStock

ReadiLink™ Cy5 Nick Translation dsDNA Labelling Kit provides a simple and efficient way to label a double stranded DNA sample with the bright and photostable Cy5 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 Cy5 dye is conjugated. In addition, the kit allows the user to optimize incorporation and product size by adjusting the ratio of Cy5-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 Cy5-labeled DNAs can be used in a variety of molecular biology techniques such as fluorescence in situ hybridization (FISH).

Example protocol

AT A GLANCE

Protocol summary

Prepare DNA samples
Add reagents to tube
Mix and centrifuge briefly
Incubate at 15 °C for 60 minutes
Place the reaction on ice followed by addition of Stop Solution and heating at 65 °C
Place on ice for 5 minutes before using or store at 4 °C
Purify the labelled DNA

Important

Thaw all the kit components on ice before starting the experiment. Briefly vortex all the reagents to the bottom before starting the labelling process.

SAMPLE EXPERIMENTAL PROTOCOL

The following protocol can be used as a guideline.
Table 1.Reagents composition per tube for each reaction

Components	Amount
DNA sample	1 µg DNA diluted in Nuclease-free water to final volume of 34 µL
Nick Translation Buffer	5 µL
dNTP mix	5 µL
dTTP	2 µL
Cy5-dUTP working solution	2 µL
DNA Polymerase I	1 µL
DNase I	1 µL
Total Volume	50 µL

The ratio of Cy5-dUTP (Component A): dTTP (Component E) can be optimized to achieve the best labelling conditions.
Incubation time can be optimized for better labelling. Longer incubation time will help with more labelling but may shorten the size of the end product.

To a clean (Nuclease-free) 0.5 mL micro centrifuge tube or 0.2 mL PCR tube, add the reagents in the order indicated in Table 1.
Carefully mix the reagents by a brief vortex followed by brief centrifuge.
Incubate the reaction at 15 °C for 60 minutes.
After incubation, place the reaction on ice.
To terminate the reaction, add 5 µL of Stop Solution and heat the sample at 65 °C.
Place on ice for 5 minutes before using or store at 4 °C.
Purify the labeled DNA.

Spectrum

Open in Advanced Spectrum Viewer

Product family

Name	Excitation (nm)	Emission (nm)	Extinction coefficient (cm ^-1 M ^-1)	Quantum yield	Correction Factor (260 nm)	Correction Factor (280 nm)
ReadiLink™ Cy3 Nick Translation dsDNA Labeling Kit	555	569	150000¹	0.15¹	0.07	0.073

References

View all 50 references: Citation Explorer

Customized optical mapping by CRISPR-Cas9 mediated DNA labeling with multiple sgRNAs.
Authors: Abid, Heba Z and Young, Eleanor and McCaffrey, Jennifer and Raseley, Kaitlin and Varapula, Dharma and Wang, Hung-Yi and Piazza, Danielle and Mell, Joshua and Xiao, Ming
Journal: Nucleic acids research (2021): e8

Assessment of Global DNA Double-Strand End Resection using BrdU-DNA Labeling coupled with Cell Cycle Discrimination Imaging.
Authors: O'Sullivan, Julia and Mersaoui, Sofiane Y and Poirier, Guy and Masson, Jean-Yves
Journal: Journal of visualized experiments : JoVE (2021)

Coupled DNA-labeling and sequencing approach enables the detection of viable-but-non-culturable Vibrio spp. in irrigation water sources in the Chesapeake Bay watershed.
Authors: Malayil, Leena and Chattopadhyay, Suhana and Mongodin, Emmanuel F and Sapkota, Amy R
Journal: Environmental microbiome (2021): 13

Fast and Efficient Postsynthetic DNA Labeling in Cells by Means of Strain-Promoted Sydnone-Alkyne Cycloadditions.
Authors: Krell, Katja and Pfeuffer, Bastian and Rönicke, Franziska and Chinoy, Zoeisha S and Favre, Camille and Friscourt, Frédéric and Wagenknecht, Hans-Achim
Journal: Chemistry (Weinheim an der Bergstrasse, Germany) (2021)

Metabolically-active bacteria in reclaimed water and ponds revealed using bromodeoxyuridine DNA labeling coupled with 16S rRNA and shotgun sequencing.
Authors: Malayil, Leena and Ramachandran, Padmini and Chattopadhyay, Suhana and Cagle, Robin and Hittle, Lauren and Ottesen, Andrea and Mongodin, Emmanuel F and Sapkota, Amy R
Journal: Water research (2020): 116185

Page updated on January 31, 2025

Ordering information

Price
Unit size	10 Reactions 20 Reactions
Catalog Number	1740817409
Quantity

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Additional ordering information

Telephone	1-800-990-8053
Fax	1-800-609-2943
Email	sales@aatbio.com
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Purchase order	Send to sales@aatbio.com
Shipping	Standard overnight for United States, inquire for international

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Spectral properties

Correction Factor (260 nm)	0.02
Correction Factor (280 nm)	0.03
Correction Factor (482 nm)	0.009
Correction Factor (565 nm)	0.09
Extinction coefficient (cm ^-1 M ^-1)	250000¹
Excitation (nm)	651
Emission (nm)	670
Quantum yield	0.27¹, 0.4²