BIOTIN-16-DUTP SOLUTION

General description

Biotin is a water-soluble vitamin which functions as a coenzyme for carboxylases.Biotin-16-dUTP is used to produce biotinylated DNA probes that are commonly used to detect specific DNA and RNA sequences in different methods like Southern and Northern blotting, in situ hybridization, and sandwich assays.

Application

Biotin-16-dUTP has been used:

• For nonradioactive DNA labeling such as random priming, PCR labeling, or nick translation. Biotin-16-dUTP replaces dTTP in the random-primed DNA labeling reaction or nick translation in a ratio of 35% Biotin-16-dUTP and 65% dTTP, as well as in PCR.
• as a substrate for terminal transferase, DNA polymerase I (holoenzyme and Klenow fragment), Taq DNA polymerase, and reverse transcriptase (e.g., Transcriptor). The nucleotide also serves as a substrate for Terminal Transferase in 3′-end labeling
• In fluorescence in situ hybridization (FISH)

Biochem/physiol Actions

Biotin influences gene regulation and chromatin structure by associating with lysine residues in histones. It influences the actions of adaptive immune T and natural killer cells. Deficiency of biotin in mice has indicated enhanced inflammation. Studies have also indicated heightened inflammatory response of human dendritic cells in conditions of biotin deficiency.

Quality

Typical analysis: At least 85% deoxynucleoside triphosphate (Biotin-16-dUTP) (HPLC, area%).
Notes: For your convenience, biotin-dUTP is also available in convenient labeling mixes (single solution with conventional dNTPs and biotin-16-dUTP), and technique-specific mixes (nucleotides, buffers, and enzymes in a single vial).

Analysis Note

Absorption: The spectral data of Biotin-16-dUTP (at pH 7.3):
Emax₂₄₁: E = 10.700;
Emax₂₉₂: E = 7.000;
Emax₂₆₅: E = 4.400;
A_292/265 : 1.6.

Other Notes

Biotin-labeled DNA can be detected with: Streptavidin-alkaline phosphatase conjugate and a chemiluminescent substrate (CSPD, CDP-Star) or a color substrate- Biotin Luminescence Detection Kit.