Biotin-14-dCTP

For general laboratory use.

Please centrifuge briefly before opening (volume ≤2 ml).

Description:
Biotin-14-dCTP is enzymatically incorporated into DNA/cDNA as substitute for its natural counterpart dCTP. The resulting Biotin-labeled DNA/cDNA probes are subsequently detected using streptavidin conjugated with horseradish peroxidase (HRP), alkaline phosphatase (AP), a fluorescent dye or agarose/magnetic beads. Optimal substrate properties for Nick Translation are ensured by a 14-atom linker attached to the N4 position of cytosine. For PCR incorporation experiments e.g. with Taq polymerase Biotin-11-dCTP (#NU-809-BIOX) or Biotin-16-dCTP (#NU-809-BIO16) are recommended whose Biotin moiety is attached to C5 position of cytidine via a 11-atom or 16-atom linker, respectively.

Recommended Biotin-14-dCTP/dCTP ratio for Nick Translation: 50% Biotin-14-dCTP/ 50% dCTP

Please note: The optimal final concentration of Biotin-14-dCTP may very depending on the application and assay conditions. For optimal product yields and high incorporation rates an individual optimization of the Biotin-14-dCTP/dCTP ratio is recommended.

Shipping: shipped on gel packs

Storage Conditions: store at -20 °C
Short term exposure (up to 1 week cumulative) to ambient temperature possible.

Shelf Life: 12 months after date of delivery

Molecular Formula: C₃₁H₅₄N₇O₁₆P₃S (free acid)

Molecular Weight: 905.78 g/mol (free acid)

Exact Mass: 905.26 g/mol (free acid)

Purity: ≥ 95 % (HPLC)

Form: filtered solution (30 kDa) in 10 mM Tris-HCl

Color: colorless to slightly yellow

Concentration: 1.0 mM - 1.1 mM

pH: 7.5 ±0.5

Spectroscopic Properties: λ_max 272 nm, ε 13.3 L mmol^-1 cm^-1 (Tris-HCl pH 7.5)

Applications:
Incorporation into DNA/cDNA by
- Nick Translation with DNAse I/ DNA Polymerase I ^{[1] & in-house data}
- Primer Extension with Klenow fragment ^[2]

Selected References:
[1] Gebeyehu et al. (1987) Novel biotinylated nucleotide-analogs for labeling and colorimetric detection of DNA. Nucleic Acids Res. 15 (21):4513.
[2] Lieberman-Aiden et al. (2009) Comprehensive mapping of long range interactions reveals folding principles of the human genome. Science 326 (5959):289.