ReadiLink™ Cy5 Oligo and ssDNA Labeling Kit

ReadiLink™ Cy5 Oligo and ssDNA Labelling Kit enables simple and uniform tagging of single-stranded DNA or oligos with Cy5 fluorophore. The labelling kit uses our proprietary TAQuest™ terminal deoxynucleotidyl transferase (TdT) to catalyze non-template directed nucleotide incorporation onto the 3’- end of single-stranded DNAs or oligos. The kit is optimized for efficient labelling and contains all the essential reagents required for efficient labelling of ssDNA or oligos. The resulting Cy5-labelled DNA probes are ideally suited for biological applications, e.g., electrophoretic mobility shift assays (EMSA), Northern and Southern blots, colony or in situ hybridizations.

Example protocol

AT A GLANCE

Protocol summary

Prepare oligo or ssDNA samples
Add reagents to tube
Mix and centrifuge briefly
Incubate at 37 °C for 60 minutes
Place on ice for 5 minutes
Purify the labeled DNA

Note: Thaw all the kit components on ice before starting the experiment. Briefly centrifuge all the reagents to the bottom before starting the labeling process.

SAMPLE EXPERIMENTAL PROTOCOL

The following protocol can be used as a guideline.

To a clean (Nuclease-free) 0.5 mL microcentrifuge tube or 0.2 mL PCR tube, prepare a reaction mix by adding the reagents in the order indicated in Table 1.
Carefully mix the reagents by a brief vortex, followed by a brief centrifuge.
Incubate the reaction at 37 °C for 60 minutes.
After incubation, place the reaction on ice for 5 minutes.
Purify the labeled DNA.

Table 1.Reagents composition per tube for each reaction

Components	Amount
Oligo or ssDNA sample	1 µg DNA diluted in Nuclease-free water to a final volume of 5 µL
TdT Reaction Buffer	40 µL
Cy5-dUTP	1-2 µL
CoCl₂	5 µL
TdT enzyme	0.5 µL
Total Volume	52 µL (Approx.)

Note: The amount of Cy5-dUTP can be optimized to achieve the best labeling conditions.

Spectrum

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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 Oligo and ssDNA Labeling Kit	555	569	150000¹	0.15¹	0.07	0.073

References

View all 2 references: Citation Explorer

A High-Throughput Image Correlation Method for Rapid Analysis of Fluorophore Photoblinking and Photobleaching Rates.
Authors: Sehayek, Simon and Gidi, Yasser and Glembockyte, Viktorija and Brandão, Hugo B and François, Paul and Cosa, Gonzalo and Wiseman, Paul W
Journal: ACS nano (2019): 11955-11966

Base-Excision-Repair-Induced Construction of a Single Quantum-Dot-Based Sensor for Sensitive Detection of DNA Glycosylase Activity.
Authors: Wang, Li-Juan and Ma, Fei and Tang, Bo and Zhang, Chun-Yang
Journal: Analytical chemistry (2016): 7523-9

Page updated on November 21, 2024

Ordering information

Price
Unit size	10 Reactions 20 Reactions
Catalog Number	1748817489
Quantity

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Additional ordering information

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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