ReadiLink™ iFluor® 555 Oligo and ssDNA Labeling Kit

ReadiLink™ iFluor® 555 Oligo and ssDNA Labelling Kit enables simple and uniform tagging of single-stranded DNA or oligos with iFluor® 555, our bright, photostable and green-fluorescent 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 iFluor® 555-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
iFluor™ 555-dUTP	1-2 µL
CoCl₂	5 µL
TdT enzyme	0.5 µL
Total Volume	52 µL (Approx.)

Note: The amount of iFluor™ 555-dUTP can be optimized to achieve the best labeling conditions.

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)	Correction Factor (656 nm)
ReadiLink™ iFluor® 488 Oligo and ssDNA Labeling Kit	491	516	75000¹	0.9¹	0.21	0.11	-
ReadiLink™ iFluor® 647 Oligo and ssDNA Labeling Kit	656	670	250000¹	0.25¹	0.03	0.03	0.0793

References

View all 7 references: Citation Explorer

Time-Gated Luminescent In Situ Hybridization (LISH): Highly Sensitive Detection of Pathogenic Staphylococcus aureus.
Authors: Sayyadi, Nima and Connally, Russell E and Lawson, Thomas S and Yuan, Jingli and Packer, Nicolle H and Piper, James A
Journal: Molecules (Basel, Switzerland) (2019)

Movement of a Quantum Dot Covered with Cytocompatible and pH-Responsible Phospholipid Polymer Chains under a Cellular Environment.
Authors: Liu, Yihua and Oda, Haruka and Inoue, Yuuki and Ishihara, Kazuhiko
Journal: Biomacromolecules (2016): 3986-3994

Photosensitizer and polycationic peptide-labeled streptavidin as a nano-carrier for light-controlled protein transduction.
Authors: Minamihata, Kosuke and Maeda, Yasukazu and Yamaguchi, Satoshi and Ishihara, Wataru and Ishiwatari, Akira and Takamori, Satoshi and Yamahira, Shinya and Nagamune, Teruyuki
Journal: Journal of bioscience and bioengineering (2015): 630-6

Fabrication of reversible poly(dimethylsiloxane) surfaces via host-guest chemistry and their repeated utilization in cardiac biomarker analysis.
Authors: Zhang, Yanrong and Ren, Li and Tu, Qin and Wang, Xueqin and Liu, Rui and Li, Li and Wang, Jian-Chun and Liu, Wenming and Xu, Juan and Wang, Jinyi
Journal: Analytical chemistry (2011): 9651-9

Multifunctional surface modification of gold-stabilized nanoparticles by bioorthogonal reactions.
Authors: Li, Xiuru and Guo, Jun and Asong, Jinkeng and Wolfert, Margreet A and Boons, Geert-Jan
Journal: Journal of the American Chemical Society (2011): 11147-53

Page updated on November 20, 2024

Ordering information

Price
Unit size	10 Reactions 20 Reactions
Catalog Number	1748217483
Quantity

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Additional ordering information

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Fax	1-800-609-2943
Email	sales@aatbio.com
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Spectral properties

Correction Factor (260 nm)	0.23
Correction Factor (280 nm)	0.14
Extinction coefficient (cm ^-1 M ^-1)	100000¹
Excitation (nm)	557
Emission (nm)	570
Quantum yield	0.64¹