- Biotin-16-ddUTP is used as a substrate for: Terminal Transferase
- DNA polymerase I (Holoenzyme and Klenow fragment)
- T4 DNA polymerase
- Taq DNA polymerase and reverse transcriptase (e.g.,Transcriptor).
Biotin-16-ddUTP is preferentially used for 3μ-end labeling of oligonucleotides with Terminal Transferase, recombinant and labeling DNA.
- The biotin-labeled oligonucleotide can be used as a hybridization probe for: DNA and RNA transfers
- Colony and plaque screening
- In situ hybridization
In addition to common hybridization techniques, biotin-labeled oligonucleotides are specifically useful for screening expression libraries for sequence-specific DNA-binding proteins, for example, transcription factors.
The labeled oligomer can be subsequently detected by ELISA using the Streptavidin-AP conjugate for nucleic acid detection.
Oligonucleotides are enzymatically labeled at their 3μ end with Terminal Transferase by incorporation of a single biotin-labeled dideoxyuridine- triphosphate (biotin-ddUTP).
It has been used in the biotinylation of in vitro transcribed tmRNA during surface plasmon resonance (SPR) technique. It has been used in the standard nick translation reaction for tumor DNA labelling during comparative genomic hybridization.
