Application

For 3′-end labeling of oligonucleotides from 14 to 100 nucleotides in length with DIG-11-ddUTP and recombinant Terminal Transferase. DIG-labeled oligonucleotides are used in a variety of hybridization techniques: ??Dot/slot blots
??Colony/ plaque hybridizations
??Southern 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

General description

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Kit for 3′-end labeling of oligonucleotides. In this method, the enzyme terminal transferase catalyzes the addition of single digoxigenin-labeled dideoxyuridine triphosphates to the 3′-OH end of oligonucleotides to be labeled.

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 9 components

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

Quality

Function tested in a dot blot.