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ReadiLink™ Cy3 Oligo and ssDNA Labeling Kit

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Spectral properties
Correction Factor (260 nm)0.07
Correction Factor (280 nm)0.073
Extinction coefficient (cm -1 M -1)1500001
Excitation (nm)555
Emission (nm)569
Quantum yield0.151
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
UNSPSC12171501
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OverviewpdfSDSpdfProtocol


Correction Factor (260 nm)
0.07
Correction Factor (280 nm)
0.073
Extinction coefficient (cm -1 M -1)
1500001
Excitation (nm)
555
Emission (nm)
569
Quantum yield
0.151
ReadiLink™ Cy3 Oligo and ssDNA Labelling Kit enables simple and uniform tagging of single-stranded DNA or oligos with Cy3 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 Cy3-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.

Platform


Thermal Cycler

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

Components


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
ComponentsAmount
Oligo or ssDNA sample1 µg DNA diluted in Nuclease-free water to a final volume of 5 µL
TdT Reaction Buffer40 µL
Cy3-dUTP1-2 µL
CoCl25 µL
TdT enzyme0.5 µL
Total Volume52 µL (Approx.)
Note: The amount of Cy3-dUTP can be optimized to achieve the best labeling conditions.

Spectrum


Open in Advanced Spectrum Viewer
spectrum

Spectral properties

Correction Factor (260 nm)0.07
Correction Factor (280 nm)0.073
Extinction coefficient (cm -1 M -1)1500001
Excitation (nm)555
Emission (nm)569
Quantum yield0.151

Product Family


NameExcitation (nm)Emission (nm)Extinction coefficient (cm -1 M -1)Quantum yieldCorrection Factor (260 nm)Correction Factor (280 nm)Correction Factor (482 nm)Correction Factor (565 nm)
ReadiLink™ Cy5 Oligo and ssDNA Labeling Kit65167025000010.271, 0.420.020.030.0090.09

References


View all 50 references: Citation Explorer
Frequency-Upconverted Stimulated Emission by Up to Six-Photon Excitation from Highly Extended Spiro-Fused Ladder-Type Oligo(p-phenylene)s.
Authors: Jiang, Yi and Li, King Fai and Gao, Kun and Lin, He and Tam, Hoi Lam and Liu, Yuan-Yuan and Shu, Yu and Wong, Ka-Leung and Lai, Wen-Yong and Cheah, Kok Wai and Huang, Wei
Journal: Angewandte Chemie (International ed. in English) (2021): 10007-10015
Oligo-FISH barcode in beans: a new chromosome identification system.
Authors: de Oliveira Bustamante, Fernanda and do Nascimento, Thiago Henrique and Montenegro, Claudio and Dias, Sibelle and do Vale Martins, Lívia and Braz, Guilherme Tomaz and Benko-Iseppon, Ana Maria and Jiang, Jiming and Pedrosa-Harand, Andrea and Brasileiro-Vidal, Ana Christina
Journal: TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik (2021): 3675-3686
Distribution of FISH oligo-5S rDNA and oligo-(AGGGTTT)3 in Hibiscus mutabilis L.
Authors: Luo, Xiaomei and He, Zhoujian
Journal: Genome (2021): 655-664
Identification of 5S and 45S rDNA sites in Chrysanthemum species by using oligonucleotide fluorescence in situ hybridization (Oligo-FISH).
Authors: He, Jun and Lin, Sisi and Yu, Zhongyu and Song, Aiping and Guan, Zhiyong and Fang, Weimin and Chen, Sumei and Zhang, Fei and Jiang, Jiafu and Chen, Fadi and Wang, Haibin
Journal: Molecular biology reports (2021): 21-31
Bottom-up modular synthesis of well-defined oligo(arylfuran)s.
Authors: Chen, Yang and Shen, Pingchuan and Cao, Tongxiang and Chen, Hao and Zhao, Zujin and Zhu, Shifa
Journal: Nature communications (2021): 6165
Triazolyl Conjugated (Oligo)Phenothiazines Building Blocks for Hybrid Materials-Synthesis and Electronic Properties.
Authors: Khelwati, Hilla and Franz, Adam W and Zhou, Zhou and Thiel, Werner R and Müller, Thomas J J
Journal: Molecules (Basel, Switzerland) (2021)
Understanding and Tailoring Excited State Properties in Solution-Processable Oligo(p-phenyleneethynylene)s: Highly Fluorescent Hybridized Local and Charge Transfer Character via Experiment and Theory.
Authors: Usta, Hakan and Cosut, Bunyemin and Alkan, Fahri
Journal: The journal of physical chemistry. B (2021): 11717-11731
Chromosomal Characterization of Tripidium arundinaceum Revealed by Oligo-FISH.
Authors: Yu, Fan and Chai, Jin and Li, Xueting and Yu, Zehuai and Yang, Ruiting and Ding, Xueer and Wang, Qiusong and Wu, Jiayun and Yang, Xiping and Deng, Zuhu
Journal: International journal of molecular sciences (2021)
An efficient Oligo-FISH painting system for revealing chromosome rearrangements and polyploidization in Triticeae.
Authors: Li, Guangrong and Zhang, Tao and Yu, Zhihui and Wang, Hongjin and Yang, Ennian and Yang, Zujun
Journal: The Plant journal : for cell and molecular biology (2021): 978-993
The Puzzling Fate of a Lupin Chromosome Revealed by Reciprocal Oligo-FISH and BAC-FISH Mapping.
Authors: Bielski, Wojciech and Książkiewicz, Michał and Šimoníková, Denisa and Hřibová, Eva and Susek, Karolina and Naganowska, Barbara
Journal: Genes (2020)