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ReadiLink™ Biotin Oligo and ssDNA Labeling Kit
ReadiLink™ Biotin Oligo and ssDNA Labelling Kit enables simple and uniform tagging of single-stranded DNA or oligos with biotin tag. 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 biotinylated oligo and ssDNA probes are ideally suited for a variety of 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 Biotin-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 |
| Biotin-dUTP | 1-2 µL |
| CoCl2 | 5 µL |
| TdT enzyme | 0.5 µL |
| Total Volume | 52 µL (Approx.) |
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 (482 nm) | Correction Factor (565 nm) |
| ReadiLink™ Cy3 Oligo and ssDNA Labeling Kit | 555 | 569 | 1500001 | 0.151 | 0.07 | 0.073 | - | - |
| ReadiLink™ Cy5 Oligo and ssDNA Labeling Kit | 651 | 670 | 2500001 | 0.271, 0.42 | 0.02 | 0.03 | 0.009 | 0.09 |
References
View all 36 references: Citation Explorer
Electrofluorochromic Imaging Analysis of Glycan Expression on Living Single Cell with Bipolar Electrode Arrays.
Authors: Tian, Zhaoyan and Wu, Yafeng and Shao, Fengying and Tang, Dezhi and Qin, Xiang and Wang, Chenchen and Liu, Songqin
Journal: Analytical chemistry (2021): 5114-5122
Authors: Tian, Zhaoyan and Wu, Yafeng and Shao, Fengying and Tang, Dezhi and Qin, Xiang and Wang, Chenchen and Liu, Songqin
Journal: Analytical chemistry (2021): 5114-5122
Duplex-specific nuclease-based electrochemical biosensor for the detection of microRNAs by conversion of homogeneous assay into surface-tethered electrochemical analysis.
Authors: Liu, Lin and Deng, Dehua and Wu, Daohong and Hou, Weilin and Wang, Lu and Li, Ning and Sun, Zhifang
Journal: Analytica chimica acta (2021): 338199
Authors: Liu, Lin and Deng, Dehua and Wu, Daohong and Hou, Weilin and Wang, Lu and Li, Ning and Sun, Zhifang
Journal: Analytica chimica acta (2021): 338199
An aggregation-induced emission fluorogen/DNA probe carrying an endosome escaping pass for tracking reduced thiol compounds in cells.
Authors: Hu, Yinhua and Cao, Xiuping and Guo, Yingshu and Zheng, Xiaofei and Li, Dongjiao and Chen, Si-Kai and Chen, Guang and You, Jinmao
Journal: Analytical and bioanalytical chemistry (2020): 7811-7817
Authors: Hu, Yinhua and Cao, Xiuping and Guo, Yingshu and Zheng, Xiaofei and Li, Dongjiao and Chen, Si-Kai and Chen, Guang and You, Jinmao
Journal: Analytical and bioanalytical chemistry (2020): 7811-7817
A voltammetric biosensor for mercury(II) using reduced graphene oxide@gold nanorods and thymine-Hg(II)-thymine interaction.
Authors: Jin, Huali and Zhang, Mingli and Wei, Min and Cheng, Jun-Hu
Journal: Mikrochimica acta (2019): 264
Authors: Jin, Huali and Zhang, Mingli and Wei, Min and Cheng, Jun-Hu
Journal: Mikrochimica acta (2019): 264
Metal-enhanced chemiluminescence detection of C-reaction protein based on silver nanoparticle hybrid probes.
Authors: Zong, Chen and Zhang, Duoduo and Jiang, Fan and Yang, Hua and Liu, Shijia and Li, Ping
Journal: Talanta (2019): 164-169
Authors: Zong, Chen and Zhang, Duoduo and Jiang, Fan and Yang, Hua and Liu, Shijia and Li, Ping
Journal: Talanta (2019): 164-169
Page updated on November 21, 2024
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