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Roche

DIG Oligonucleotide 3′-End Labeling Kit, 2nd generation

greener alternative

sufficient for 25 labeling reactions (100 pmol of oligonucleotides per assay; 1 ug of a 30-mer oligonucleotide), storage condition avoid repeated freeze/thaw cycles

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About This Item

UNSPSC Code:
12352200

usage

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

Quality Level

manufacturer/tradename

Roche

greener alternative product characteristics

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

greener alternative category

shipped in

dry ice

General description

The DIG Oligonucleotide 3′-End Labeling Kit, 2nd generation employs the enzyme terminal transferase. It catalyzes the addition of single digoxigenin-labeled dideoxyuridine triphosphates (ddUTP) to the 3′-OH end of oligonucleotides. Thus, helps in labeling 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 3′-End Labeling Kit, 2nd generation is for 3′-end labeling of oligonucleotides from 14 to 100 nucleotides in length with DIG-11-ddUTP and recombinant terminal transferase.
DIG-labeled oligonucleotides has been used in a variety of hybridization techniques:
  • dot/slot blots
  • colony/ plaque hybridizations
  • Southern blots/ northern blots
  • in situ hybridizations

Features and Benefits

  • 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, the oligonucleotides readily diffuse into fixed tissues and cells

Packaging

1 kit containing 9 components

Quality

Function tested in a dot blot.

Principle

One DIG-ddUTP molecule is added to the 3′-end of oligonucleotides by recombinant Terminal Transferase. This guarantees a very specific and distinct hybridization signal, which is detected by an enzyme-linked immunoassay with anti-DIG-AP antibody conjugate, and a color or chemiluminescence reaction.

Preparation Note

Activator: sodium sulfate, Tris
Working concentration: Oligonucleotides: 100 pmol
Up to 100 pmol (1 μg of a 30-mer) oligonucleotide can be labeled in a single standard labeling reaction.
Assay Time: The complete procedure from labeling the oligonucleotide to hybridization and detection of the first visible signal can be accomplished within less than 24 hours.

Sample Materials
Oligonucleotides of a length from 14 to 100 nucleotides, purified by HPLC or gel electrophoresis

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-ddUTP Solution 1 mM

  • Recombinant Terminal Transferase 400 U/μl

  • Control Oligonucleotide, unlabeled 20 pmol/μl

  • Oligonucleotide, DIG-ddUTP labeled 2.5 pmol/μl

  • Control DNA, 2.5 pmol/μl pUC 18 DNA, supercoiled

  • Glycogen Solution 20 mg/ml

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

See All (9)

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


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Proceedings of the National Academy of Sciences of the United States of America, 102(47), 16967-16972 (2005-11-15)
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Omer Choresh et al.
Biomacromolecules, 10(10), 2852-2856 (2009-09-08)
The various silks that make up the web of the orb web spiders have been studied extensively. However, success in prey capture depends as much on the web glue as on the fibers. Spider silk glue, which is considered one

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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