Description
General description
The DIG Oligonucleotide 3′-End Labeling Kit, 2nd generation employs the enzyme terminal transferase. It catalyzes the addition of single digoxigenin-labeled dideoxyuridine triphosphates (ddUTP) to the 3′-OH end of oligonucleotides. Thus, helps in labeling 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 3′-End Labeling Kit, 2nd generation is for 3′-end labeling of oligonucleotides from 14 to 100 nucleotides in length with DIG-11-ddUTP and recombinant terminal transferase.
DIG-labeled oligonucleotides has been used in a variety of hybridization techniques:
dot/slot blots
colony/ plaque hybridizations
Southern blots/ northern blots
in situ hybridizations
Biochem/physiol Actions
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.
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
Packaging
1 kit containing 9 components
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
Store at -15–-25 °C. (unopened kit)
Analysis Note
Function tested in a dot blot.
Other Notes
For life science research only. Not for use in diagnostic procedures.
