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Roche

DIG Oligonucleotide Tailing Kit, 2nd generation

greener alternative

sufficient for 25 reactions (100 pmol oligonucleotide per assay; 1 ug of a 30-mer oligonucleotide)

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About This Item

UNSPSC Code:
41105500
NACRES:
NA.55

usage

sufficient for 25 reactions (100 pmol oligonucleotide per assay; 1 ug of a 30-mer oligonucleotide)

Quality Level

manufacturer/tradename

Roche

storage condition

avoid repeated freeze/thaw cycles

greener alternative product characteristics

Designing Safer Chemicals
Learn more about the Principles of Green Chemistry.

sustainability

Greener Alternative Product

greener alternative category

General description

DIG Oligonucleotide Tailing Kit, 2nd generation employs the enzyme terminal transferase. It catalyzes the addition of digoxigenin (DIG)-11-deoxyuridine triphosphate (dUTP) and deoxyadenosine triphosphate (dATP) to the 3′-OH end of oligonucleotides.
We are committed to bringing you Greener Alternative Products, which adhere to one or more of The 12 Principles of Greener Chemistry. This product is designed as a safer chemical.  The DIG System was established as a sensitive and cost-effective alternative to using radioactivity for the labeling and detection of nucleic acids. There are many available publications that prove the versatility of the DIG System, so use of radio-labeling is no longer the only option for labeling of DNA for hybridization.

Application

DIG Oligonucleotide Tailing Kit, 2nd generation has been used to label oligonucleotide probes in:
  • northern blot assay
  • in situ hybridization (ISH)
  • fluorescence in situ hybridization (FISH)
In addition to common hybridization techniques, DIG-labeled oligonucleotides are especially useful for screening expression libraries for sequence-specific DNA binding proteins, for example, transcription factors.

Features and Benefits

Tailing of oligonucleotides at the 3′-end with DIG-11-dUTP and recombinant Terminal Transferase. Oligonucleotides are tailed with DIG-dUTP and dATP at an average tail length of 50 nucleotides (tail length range: 10 – 100).

  • Very sensitive hybridization probes, due to the incorporation of several DIG-nucleotides
  • Fast hybridization kinetics, due to the small size of oligonucleotides
  • Single-stranded probes, no renaturation during hybridization
  • Sequence can be designed according to the experiment
  • Specially suited for in situ hybridization; due to their small size, oligonucleotides readily diffuse into fixed tissues and cells

Packaging

1 kit containing 11 components

Principle

DIG-dUTP and dATP are combined at a concentration that gives the highest DIG incorporation into the tail, and optimal spacing of DIG and dATP, to achieve the highest sensitivity in hybridization experiments. DIG-dUTP and dATP are provided as separate solutions to allow greater flexibility in terms of tail length, hapten spacing, and the use of unlabeled nucleotide(s).

Preparation Note

Working concentration: Oligonucleotides
In one standard labeling reaction up to 100 pmol oligonucleotide (1 μg of a 30-mer oligonucleotide) can be applied.

Storage and Stability

Store at -15–-25 °C. (unopened kit)

Other Notes

For life science research only. Not for use in diagnostic procedures.

Kit Components Only

Product No.
Description

  • Reaction Buffer 5x concentrated

  • CoCl<SUB>2</SUB> Solution 25 mM

  • DIG-dUTP Solution 1 mM

  • dATP Solution 10 mM

  • Recombinant Terminal Transferase 400 U/μl

  • Control Oligonucleotide, unlabeled 20 pmol/μl

  • Oligonucleotide, DIG-dUTP/dATP tailed 2.5 pmol/μl

  • Control DNA 0.25 mg/ml

  • Glycogen Solution 20 mg/ml

  • DNA Dilution Buffer, 50 μg/ml fish sperm DNA

  • Poly(A) Solution 10 mg/ml

See All (11)

signalword

Danger

Hazard Classifications

Acute Tox. 4 Inhalation - Acute Tox. 4 Oral - Aquatic Chronic 2 - Carc. 1B Inhalation - Repr. 1B

Storage Class

6.1D - Non-combustible, acute toxic Cat.3 / toxic hazardous materials or hazardous materials causing chronic effects

wgk_germany

WGK 3

flash_point_f

does not flash

flash_point_c

does not flash


Certificates of Analysis (COA)

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Xiaoming Li et al.
Nucleic acids research, 32(3), 867-877 (2004-02-13)
We report here the biochemical characterization of the deafness-associated mitochondrial tRNA(Ser(UCN)) T7511C mutation, in conjunction with homoplasmic ND1 T3308C and tRNA(Ala) T5655C mutations using cybrids constructed by transferring mitochondria from lymphoblastoid cell lines derived from an African family into human
Geeta Palsule et al.
Nucleic acids research, 47(16), 8746-8754 (2019-07-10)
RNase P RNA (RPR), the catalytic subunit of the essential RNase P ribonucleoprotein, removes the 5' leader from precursor tRNAs. The ancestral eukaryotic RPR is a Pol III transcript generated with mature termini. In the branch of the arthropod lineage
Kaposi's sarcoma-associated herpesvirus mRNA accumulation in nuclear foci is influenced by viral DNA replication and viral noncoding polyadenylated nuclear RNA
<BIG>Vallery TK, et al.</BIG>
Journal of Virology, 92, e00220-e00218 (2018)
Xian Wu Cheng et al.
The American journal of pathology, 173(2), 358-369 (2008-06-28)
The elastolytic activity of cathepsins in the myocardium is implicated in hypertensive heart failure (HF). Given that reactive oxygen species are also implicated in protease activation associated with cardiac remodeling, we examined the role of the reactive oxygen species-induced cathepsin
L1CAM in the early enteric and urogenital system
<BIG>Pechriggl E, et al.</BIG>
The Journal of Histochemistry and Cytochemistry, 65, 21-32 (2017)

Articles

Digoxigenin (DIG) labeling methods and kits for DNA and RNA DIG probes, random primed DNA labeling, nick translation labeling, 5’ and 3’ oligonucleotide end-labeling.

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