Application

This kit is used for tailing of oligonucleotides with digoxigenin-dUTP. DIG-labeled oligonucleotides are used in a variety of hybridization techniques: ??Dot/slot blot
??Colony/plaque hybridizations
??Northern blots (for abundant messages)
??Southern blots (not recommended for single-copy gene detection in complex genomes)
??In situ?hybridizations
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?

General description

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Kit for tailing of oligonucleotides at the 3′ end. In this method, the recombinant enzyme terminal transferase catalyzes the addition of DIG-11-dUTP and dATP to the 3′-OH end of oligonucleotides.

Sigma Life Science is 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.

Other Notes

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

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.