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ReadiLink™ Biotin Oligo and ssDNA Labeling Kit

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Catalog Number17490
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Telephone1-408-733-1055
Fax1-408-733-1304
Emailsales@aatbio.com
InternationalSee distributors
ShippingStandard overnight for United States, inquire for international
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


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.

Platform


Thermal Cycler

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

Components


Component A: Biotin-dUTP1 vial (20 µL)
Component B: TdT enzyme1 vial (5 µL)
Component C: CoCl2 solution1 vial (50 µL)
Component D: TdT Reaction Buffer1 vial (500 µL)

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
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.)
Note: The amount of Biotin-dUTP can be optimized to achieve the best labeling conditions.

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™ Cy3 Oligo and ssDNA Labeling Kit55556915000010.1510.070.073--
ReadiLink™ Cy5 Oligo and ssDNA Labeling Kit65167025000010.271, 0.420.020.030.0090.09

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
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
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
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
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
Graphene oxide-assisted Au nanoparticle strip biosensor based on GR-5 DNAzyme for rapid lead ion detection.
Authors: Wang, Hai-Bo and Ma, Li-Hong and Fang, Bi-Yun and Zhao, Yuan-Di and Hu, Xue-Bin
Journal: Colloids and surfaces. B, Biointerfaces (2018): 305-312
Graphene oxide@gold nanorods-based multiple-assisted electrochemiluminescence signal amplification strategy for sensitive detection of prostate specific antigen.
Authors: Cao, Jun-Tao and Yang, Jiu-Jun and Zhao, Li-Zhen and Wang, Yu-Ling and Wang, Hui and Liu, Yan-Ming and Ma, Shu-Hui
Journal: Biosensors & bioelectronics (2018): 92-98
Au nanoparticles on two-dimensional MoS2 nanosheets as a photoanode for efficient photoelectrochemical miRNA detection.
Authors: Fu, Nina and Hu, Yue and Shi, Saihua and Ren, Shaokang and Liu, Wei and Su, Shao and Zhao, Baomin and Weng, Lixing and Wang, Lianhui
Journal: The Analyst (2018): 1705-1712
Esrrb directly binds to Gata6 promoter and regulates its expression with Dax1 and Ncoa3.
Authors: Uranishi, Kousuke and Akagi, Tadayuki and Koide, Hiroshi and Yokota, Takashi
Journal: Biochemical and biophysical research communications (2016): 1720-5
Quantitative Correlation of in Vivo Properties with in Vitro Assay Results: The in Vitro Binding of a Biotin-DNA Analogue Modifier with Streptavidin Predicts the in Vivo Avidin-Induced Clearability of the Analogue-Modified Antibody.
Authors: Dou, Shuping and Virostko, John and Greiner, Dale L and Powers, Alvin C and Liu, Guozheng
Journal: Molecular pharmaceutics (2015): 3097-103