ReadiLink™ Cy5 Oligo and ssDNA Labeling Kit
ReadiLink™ Cy5 Oligo and ssDNA Labelling Kit enables simple and uniform tagging of single-stranded DNA or oligos with Cy5 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 Cy5-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 Cy5-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 |
Cy5-dUTP | 1-2 µL |
CoCl2 | 5 µL |
TdT enzyme | 0.5 µL |
Total Volume | 52 µL (Approx.) |
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) |
ReadiLink™ Cy3 Oligo and ssDNA Labeling Kit | 555 | 569 | 1500001 | 0.151 | 0.07 | 0.073 |
References
View all 2 references: Citation Explorer
A High-Throughput Image Correlation Method for Rapid Analysis of Fluorophore Photoblinking and Photobleaching Rates.
Authors: Sehayek, Simon and Gidi, Yasser and Glembockyte, Viktorija and Brandão, Hugo B and François, Paul and Cosa, Gonzalo and Wiseman, Paul W
Journal: ACS nano (2019): 11955-11966
Authors: Sehayek, Simon and Gidi, Yasser and Glembockyte, Viktorija and Brandão, Hugo B and François, Paul and Cosa, Gonzalo and Wiseman, Paul W
Journal: ACS nano (2019): 11955-11966
Base-Excision-Repair-Induced Construction of a Single Quantum-Dot-Based Sensor for Sensitive Detection of DNA Glycosylase Activity.
Authors: Wang, Li-Juan and Ma, Fei and Tang, Bo and Zhang, Chun-Yang
Journal: Analytical chemistry (2016): 7523-9
Authors: Wang, Li-Juan and Ma, Fei and Tang, Bo and Zhang, Chun-Yang
Journal: Analytical chemistry (2016): 7523-9
Page updated on December 6, 2024