The procedure for labeling of DNA probes with a polynucleotide "tail" containing hapten-labeled nucleotides was developed by Enzo.
In such terminal labeling reactions, terminal transferase catalyzes the addition of nucleotides to any 3'-OH terminus in a template independent manner. This rapid and convenient nonradioactive labeling procedure is free of any sequence bias that is normally observed in random priming or nick translation reactions. Terminal labeling is an ideal procedure for the labeling of oligonucleotides. An oligonucleotide can be synthesized using standard, commercially available reagents and labeled after synthesis in a simple and reproducible enzyme reaction. The BioProbe® 3'-Oligonucleotide Labeling System provides maximum flexibility for labeling oligonucleotides approximately 15 to 100 nucleotides in length. A standard reaction will label up to 100 picomoles (1µg of a 30-mer oligonucleotide sequence). A complete system combines a standard Reagent Pack (buffers and enzyme) with a Nucleotide Pack. All components contain sufficient reagents for 25 labeling reactions.
The system is available in two modular formats:3'-Oligonucleotide Tailing (with a "tail" of nucleotides)3'-Oligonucleotide Labeling (with a single nucleotide)The BioProbe® 3'-Oligonucleotide Labeling System Reagent Pack contains all reagents required for labeling except nucleotides. For optimun results, the reagents contained in this Pack should be used with the appropriate Nucleotide Pack.
Ordering information: Contents: 5X Reaction Buffer, 100 μl Potassium cacodylate buffer, pH 7, containing β-mercaptoethanol 10X CoCl2 Solution, 50 μl 10mM solution Terminal Deoxynucleotide Transferase, 50 μl 30 units/μl in storage buffer Control Oligonucleotide (unlabeled), 25 μl 0.2 μg/μl in TE Buffer (30-mer, 5’-TTG GGT AAC GCC AGG GTT TTC CCA GTC ACG-3’, homologous to the lac Z’ region of pUC-type vector DNAs)Control Target DNA, 25 μl 0.2 μg/μl in TE Buffer (Restriction enzyme digest of a pUC-type vector containing an Epstein-Barr Virus DNA insert)
