DIG OLIGO TAILING KIT 2ND GENERATION

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.