DIG Oligonucleotide Tailing Kit, 2nd generation

Description

• 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.
• Application: DIG Oligonucleotide Tailing Kit, 2nd generation has been used to label oligonucleotide probes in:northern blot assay[1]in situ hybridization (ISH)[2]fluorescence in situ hybridization (FISH)[3]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-nucleotidesFast hybridization kinetics, due to the small size of oligonucleotidesSingle-stranded probes, no renaturation during hybridizationSequence can be designed according to the experimentSpecially 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: OligonucleotidesIn 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.