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ReadiLink™ DIG (Digoxigenin) Oligo and ssDNA Labeling Kit

Verification of single stranded DNA labeling with ReadiLink™ DIG (Digoxigenin) Oligo and ssDNA Labeling Kit. Starting material without DIG labeling (-) and starting material with DIG labeling (+) were run on 4% agarose gel followed by gel staining with SYBR™ Gold Nucleic Acid Gel Stain. DIG labeled ssDNA has more bases due to labeling, thus  shows higher band pattern compared to control.
Verification of single stranded DNA labeling with ReadiLink™ DIG (Digoxigenin) Oligo and ssDNA Labeling Kit. Starting material without DIG labeling (-) and starting material with DIG labeling (+) were run on 4% agarose gel followed by gel staining with SYBR™ Gold Nucleic Acid Gel Stain. DIG labeled ssDNA has more bases due to labeling, thus  shows higher band pattern compared to control.
Verification of single stranded DNA labeling with ReadiLink™ DIG (Digoxigenin) Oligo and ssDNA Labeling Kit. Starting material without DIG labeling (-) and starting material with DIG labeling (+) were run on 4% agarose gel followed by gel staining with SYBR™ Gold Nucleic Acid Gel Stain. DIG labeled ssDNA has more bases due to labeling, thus  shows higher band pattern compared to control.
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Telephone1-800-990-8053
Fax1-800-609-2943
Emailsales@aatbio.com
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ShippingStandard overnight for United States, inquire for international
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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™ DIG (Digoxigenin) Oligo and ssDNA Labelling Kit enables simple and uniform tagging of single-stranded DNA or oligos with DIG 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 DIG-labeled 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


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
DIG-dUTP1-2 µL
CoCl25 µL
TdT enzyme0.5 µL
Total Volume52 µL (Approx.)
Note: The amount of DIG-dUTP can be optimized to achieve the best labeling conditions.

Images


Citations


View all 1 citations: Citation Explorer
LncRNA AL592284. 1 facilitates proliferation and metastasis of cervical cancer cells via miR-30a-5p/Vimentin/EMT axis
Authors: Zhang, Jing and Liu, Hong-li and Liu, Jing-bo and Zhang, Yuan and Liu, Jing and Li, Yan-hua
Journal: Biochemical and Biophysical Research Communications (2021): 95--102

References


View all 39 references: Citation Explorer
A Comprehensive Analysis of Northern versus Liquid Hybridization Assays for mRNAs, Small RNAs, and miRNAs Using a Non-Radiolabeled Approach.
Authors: Ahmad, Waqar and Gull, Bushra and Baby, Jasmin and Mustafa, Farah
Journal: Current issues in molecular biology (2021): 457-484
Chromosome-Specific Painting in Cucumis Species Using Bulked Oligonucleotides.
Authors: Han, Yonghua and Zhang, Tao and Thammapichai, Paradee and Weng, Yiqun and Jiang, Jiming
Journal: Genetics (2015): 771-9
Application of reverse dot blot hybridization to simultaneous detection and identification of harmful algae.
Authors: Chen, Guo Fu and Zhang, Chun Yun and Wang, Yuan Yuan and Chen, Wen
Journal: Environmental science and pollution research international (2015): 10516-28
Superresolution imaging of single DNA molecules using stochastic photoblinking of minor groove and intercalating dyes.
Authors: Miller, Helen and Zhou, Zhaokun and Wollman, Adam J M and Leake, Mark C
Journal: Methods (San Diego, Calif.) (2015): 81-8
Oligo-DNA custom macroarray for monitoring major pathogenic and non-pathogenic fungi and bacteria in the phyllosphere of apple trees.
Authors: He, Ying-Hong and Isono, Sayaka and Shibuya, Makoto and Tsuji, Masaharu and Adkar Purushothama, Charith-Raj and Tanaka, Kazuaki and Sano, Teruo
Journal: PloS one (2012): e34249
In situ hybridization detection of microRNAs.
Authors: Song, Rui and Ro, Seungil and Yan, Wei
Journal: Methods in molecular biology (Clifton, N.J.) (2010): 287-94
Feasible and reliable quantification of mRNA in Arabidopsis thaliana using optical thin-film biosensor chips.
Authors: Bai, Sulan and Wang, Fan and Zhang, Zhen and Li, Shucheng and Zhang, Jie and Zhang, Yaochuan
Journal: Journal of genetics and genomics = Yi chuan xue bao (2010): 341-6
Dual-allele dipstick assay for genotyping single nucleotide polymorphisms by primer extension reaction.
Authors: Konstantou, Jessica K and Ioannou, Penelope C and Christopoulos, Theodore K
Journal: European journal of human genetics : EJHG (2009): 105-11
Genotyping of single nucleotide polymorphisms by primer extension reaction and a dual-analyte bio/chemiluminometric assay.
Authors: Konstantou, Jessica and Ioannou, Penelope C and Christopoulos, Theodore K
Journal: Analytical and bioanalytical chemistry (2007): 1747-54
Nonisotopic detection of microRNA using digoxigenin labeled RNA probes.
Authors: Ramkissoon, Shakti H and Mainwaring, Lori A and Sloand, Elaine M and Young, Neal S and Kajigaya, Sachiko
Journal: Molecular and cellular probes (2006): 1-4