Biotin-16-dUTP

For research use only.
Please centrifuge briefly before opening (volume ≤2 ml).

Description:
Biotin-16-dUTP is enzymatically incorporated into DNA/cDNA as substitute for its natural counterpart dTTP. 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 and thus labeling efficiency as well as an efficient detection of the Biotin moiety is ensured by a 16-atom linker attached to the C5 position of uridine.

Recommended Biotin-16-dUTP/dTTP ratio for PCR and Nick Translation: 50% Biotin-16-dUTP/ 50% dTTP

Please note: The optimal final concentration of Biotin-16-dUTP may very depending on the application and assay conditions. For optimal product yields and high incorporation rates an individual optimization of the Biotin-16-dUTP/dTTP 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: C₃₂H₅₂N₇O₁₈P₃S (free acid)

Molecular Weight: 947.78 g/mol (free acid)

Exact Mass: 947.23 g/mol (free acid)

Purity: ≥ 98 % (HPLC)

Form: filtered solution (30 kDa) in water

Color: colorless to slightly yellow

Concentration: 1.0 mM - 1.1 mM

pH: 7.5 ±0.5

Spectroscopic Properties: λ_max 240 nm, ε 10.7 L mmol^-1 cm^-1 (Tris-HCl pH 7.5)

Applications:
Incorporation into DNA/cDNA by
- PCR with Taq polymerase ^{[1,2] & in-house data}
- Nick Translation with DNAse I/ DNA Polymerase I ^{[3,4] & in-house data}
- Primer Extension with Klenow exo^{- [2,5]}
- 3’-End Labeling with Terminal deoxynucleotidyl Transferase (TdT) ^[6,7]
- Reverse Transcription with MMLV Reverse Transcriptase ^[8,9]
Incorporation into RNA by
- 3’-End Labeling with Terminal deoxynucleotidyl Transferase (TdT) ^[10]

Related products:

• Biotin-11-dUTP, #NU-803-BIOX
• Biotin-16-dCTP, #NU-809-BIO16
• Biotin-11-dCTP, #NU-809-BIOX
• Digoxigenin-11-dUTP, #NU-803-DIGX

Selected References:
[1] Anderson et al. (2005) Incorporation of reporter-labeled nucleotides by DNA polymerases. Biotechniques 38:257.
[2] Cross et al. (1990) The structure of a subterminal repeated sequence present on many human chromosomes.Nucleic Acids Res. 18:6649.
[3] Langer et al. (1981) Enzymatic synthesis of biotin-labeled polynucleotides: novel nucleic acid affinity probes. Proc. Natl. Sci. USA 78:6633.
[4] Brigati et al. (1983) Detection of viral genomes in cultured cells and para nembedded tissue sections using biotin-labeled hybridization probes. Virology 12632.
[5] Perry et al. (2007) Diet and the evolution of human amylase gene copy number variation. Nat Genet 39:1256.
[6] Gorczyca et al. (1993) Detection of DNA strand breaks in individual apoptotic cells by the in situ terminal deoxynucleotidyl tranferase and nick translation assays. Caner Res 63:1954.
[7] Bassell et al. (1994) Single mRNAs visualized by ultrastructural in situ hybridization are principally localized at actin lament intersections in fibroblasts. J Cell Biol 126:863.
[8] Yih et al. (2002) Changes in gene expression profiles of human fibroblasts in response to sodium arsenite treatment. Carcinogenesis 23:867.
[9] Luo et al. (2003) Designing, testing, and validating a focused stem cell microarray for characterization of neutral stem cells and progenitor cells. Stem Cells 21:575.
[10] Rosemeyer et al. (1995) Nonradioactive 3'-End Labeling of RNA molecules of different length by Terminal Deoxynucleotidyltransferase. Analytical Biochemistry 224:446.