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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
- 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
SAMPLE EXPERIMENTAL PROTOCOL
The following protocol can be used as a guideline.
Note: The amount of iFluor™ 488-dUTP can be optimized to achieve the best labeling conditions.
- 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.
| 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.) |
Spectrum
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
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
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
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 November 20, 2024
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