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ReadiLink™ iFluor® 647 Oligo and ssDNA Labeling Kit
ReadiLink™ iFluor® 647 Oligo and ssDNA Labelling Kit enables simple and uniform tagging of single-stranded DNA or oligos with iFluor® 647, 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® 647-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.
Note: The amount of iFluor™ 647-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™ 647-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) |
| ReadiLink™ iFluor® 488 Oligo and ssDNA Labeling Kit | 491 | 516 | 750001 | 0.91 | 0.21 | 0.11 |
| ReadiLink™ iFluor® 555 Oligo and ssDNA Labeling Kit | 557 | 570 | 1000001 | 0.641 | 0.23 | 0.14 |
References
View all 13 references: Citation Explorer
A Ratiometric Fluorescent Conjugated Oligomer for Amyloid β Recognition, Aggregation Inhibition, and Detoxification.
Authors: Zhang, Ziqi and Yuan, Qiong and Li, Meiqi and Bao, Benkai and Tang, Yanli
Journal: Small (Weinheim an der Bergstrasse, Germany) (2021): e2104581
Authors: Zhang, Ziqi and Yuan, Qiong and Li, Meiqi and Bao, Benkai and Tang, Yanli
Journal: Small (Weinheim an der Bergstrasse, Germany) (2021): e2104581
Chorus2: design of genome-scale oligonucleotide-based probes for fluorescence in situ hybridization.
Authors: Zhang, Tao and Liu, Guanqing and Zhao, Hainan and Braz, Guilherme T and Jiang, Jiming
Journal: Plant biotechnology journal (2021): 1967-1978
Authors: Zhang, Tao and Liu, Guanqing and Zhao, Hainan and Braz, Guilherme T and Jiang, Jiming
Journal: Plant biotechnology journal (2021): 1967-1978
General Method for Determining Redox Potentials without Electrolyte.
Authors: Bird, Matthew J and Pearson, Matthew A and Asaoka, Sadayuki and Miller, John R
Journal: The journal of physical chemistry. A (2020): 5487-5495
Authors: Bird, Matthew J and Pearson, Matthew A and Asaoka, Sadayuki and Miller, John R
Journal: The journal of physical chemistry. A (2020): 5487-5495
Breaking Parallel Orientation of Rods via a Dendritic Architecture toward Diverse Supramolecular Structures.
Authors: Zhang, Ruimeng and Feng, Xueyan and Zhang, Rui and Shan, Wenpeng and Su, Zebin and Mao, Jialin and Wesdemiotis, Chrys and Huang, Jiahao and Yan, Xiao-Yun and Liu, Tong and Li, Tao and Huang, Mingjun and Lin, Zhiwei and Shi, An-Chang and Cheng, Stephen Z D
Journal: Angewandte Chemie (International ed. in English) (2019): 11879-11885
Authors: Zhang, Ruimeng and Feng, Xueyan and Zhang, Rui and Shan, Wenpeng and Su, Zebin and Mao, Jialin and Wesdemiotis, Chrys and Huang, Jiahao and Yan, Xiao-Yun and Liu, Tong and Li, Tao and Huang, Mingjun and Lin, Zhiwei and Shi, An-Chang and Cheng, Stephen Z D
Journal: Angewandte Chemie (International ed. in English) (2019): 11879-11885
Manufacture of Clinical-Grade Human Clonal Mesenchymal Stem Cell Products from Single Colony Forming Unit-Derived Colonies Based on the Subfractionation Culturing Method.
Authors: Yi, TacGhee and Kim, Si-na and Lee, Hyun-Joo and Kim, Junghee and Cho, Yun-Kyoung and Shin, Dong-Hee and Tak, Sun-Ji and Moon, Sun-Hwa and Kang, Ji-Eun and Ji, In-Mi and Lim, Huyn-Ja and Lee, Dong-Soon and Jeon, Myung-Shin and Song, Sun U
Journal: Tissue engineering. Part C, Methods (2015): 1251-62
Authors: Yi, TacGhee and Kim, Si-na and Lee, Hyun-Joo and Kim, Junghee and Cho, Yun-Kyoung and Shin, Dong-Hee and Tak, Sun-Ji and Moon, Sun-Hwa and Kang, Ji-Eun and Ji, In-Mi and Lim, Huyn-Ja and Lee, Dong-Soon and Jeon, Myung-Shin and Song, Sun U
Journal: Tissue engineering. Part C, Methods (2015): 1251-62
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