ReadiLink™ iFluor® 488 Oligo and ssDNA Labeling Kit

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Additional ordering information

Telephone	1-800-990-8053
Fax	1-800-609-2943
Email	sales@aatbio.com
International	See distributors
Bulk request	Inquire
Custom size	Inquire
Shipping	Standard overnight for United States, inquire for international

Spectral properties

Correction Factor (260 nm)	0.21
Correction Factor (280 nm)	0.11
Extinction coefficient (cm ^-1 M ^-1)	75000¹
Excitation (nm)	491
Emission (nm)	516
Quantum yield	0.9¹

Storage, safety and handling

H-phrase	H303, H313, H333
Hazard symbol	XN
Intended use	Research Use Only (RUO)
R-phrase	R20, R21, R22
UNSPSC	12171501

Overview SDS Protocol

Correction Factor (260 nm)

0.21

Correction Factor (280 nm)

0.11

Extinction coefficient (cm ^-1 M ^-1)

75000¹

Excitation (nm)

491

Emission (nm)

516

Quantum yield

0.9¹

ReadiLink™ iFluor® 488 Oligo and ssDNA Labeling Kit enables simple and uniform tagging of single-stranded DNA or oligos with iFluor® 488, our bright, photostable and green-fluorescent fluorophore. This labeling kit uses our proprietary 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 labeling and contains all the essential reagents required for efficient labeling of ssDNA or oligos. The resulting iFluor® 488-labeled DNA probes are ideally suited for biological applications, e.g., electrophoretic mobility shift assays (EMSA), Northern and Southern blots, colony or in situ hybridizations.

Platform

Thermal Cycler

Instrument specification(s)

0.5 mL microcentrifuge or 0.2 mL PCR tube

Components

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™ 488-dUTP	1-2 µL
CoCl₂	5 µL
TdT enzyme	0.5 µL
Total Volume	52 µL (Approx.)

Note: The amount of iFluor™ 488-dUTP can be optimized to achieve the best labeling conditions.

Spectrum

Open in Advanced Spectrum Viewer

Spectral properties

Correction Factor (260 nm)	0.21
Correction Factor (280 nm)	0.11
Extinction coefficient (cm ^-1 M ^-1)	75000¹
Excitation (nm)	491
Emission (nm)	516
Quantum yield	0.9¹

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® 555 Oligo and ssDNA Labeling Kit	557	570	100000¹	0.64¹	0.23	0.14	-
ReadiLink™ iFluor® 647 Oligo and ssDNA Labeling Kit	656	670	250000¹	0.25¹	0.03	0.03	0.0793

References

View all 3 references: Citation Explorer

Two-color, 30 second microwave-accelerated Metal-Enhanced Fluorescence DNA assays: a new Rapid Catch and Signal (RCS) technology.
Authors: Dragan, Anatoliy I and Golberg, Karina and Elbaz, Amit and Marks, Robert and Zhang, Yongxia and Geddes, Chris D
Journal: Journal of immunological methods (2011): 1-7

Application of four dyes in gene expression analyses by microarrays.
Authors: Staal, Yvonne C M and van Herwijnen, Marcel H M and van Schooten, Frederik J and van Delft, Joost H M
Journal: BMC genomics (2005): 101

A fluorescence resonance energy transfer sensor for the beta-domain of metallothionein.
Authors: Hong, Sung-Hye and Maret, Wolfgang
Journal: Proceedings of the National Academy of Sciences of the United States of America (2003): 2255-60