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ReadiLink™ iFluor® 488 Oligo and ssDNA Labeling Kit

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.

Example protocol

AT A GLANCE

Protocol summary
  1. Prepare oligo or ssDNA samples
  2. Add reagents to tube
  3. Mix and centrifuge briefly
  4. Incubate at 37 °C for 60 minutes
  5. Place on ice for 5 minutes
  6. 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.
  1. 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.
  2. Carefully mix the reagents by a brief vortex, followed by a brief centrifuge.
  3. Incubate the reaction at 37 °C for 60 minutes.
  4. After incubation, place the reaction on ice for 5 minutes.
  5. 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

Product family

NameExcitation (nm)Emission (nm)Extinction coefficient (cm -1 M -1)Quantum yieldCorrection Factor (260 nm)Correction Factor (280 nm)Correction Factor (656 nm)
ReadiLink™ iFluor® 555 Oligo and ssDNA Labeling Kit55757010000010.6410.230.14-
ReadiLink™ iFluor® 647 Oligo and ssDNA Labeling Kit65667025000010.2510.030.030.0793

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
Page updated on December 6, 2024

Ordering information

Price
Unit size
10 Reactions
20 Reactions
Catalog Number
1748017481
Quantity
Add to cart

Additional ordering information

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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-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
UNSPSC12171501

Platform

Thermal Cycler

Instrument specification(s)0.5 mL microcentrifuge or 0.2 mL PCR tube

Components