Biotin-16-dCTP

For general laboratory use.

Envio: shipped on gel packs

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

Validade: 12 months after date of delivery

Fórmula molecular: C32H51N8O17P3S (free acid)

Peso molecular: 944.78 g/mol (free acid)

Pureza: ≥ 95 % (HPLC)

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

Concentração: 1.0 mM - 1.1 mM

pH: 7.5 ±0.5

Propriedades espectroscópicas: λmax 294 nm, ε 9.3 L mmol-1 cm-1 (Tris-HCl pH 7.5)

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

Descrição:
Biotin-16-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 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 cytidine.

Recommended Biotin-16-dCTP/dCTP ratio for PCR: 50% Biotin-16-dCTP/ 50% dCTP

Please note: The optimal final concentration of Biotin-16-dCTP may very depending on the application and assay conditions. For optimal product yields and high incorporation rates an individual optimization of the Biotin-16-dCTP/dTTP ratio is recommended.

Referências selecionadas:
[1] Sugimoto et al. (1999) Isolation and mapping of a putative b subunit of human ATP synthase (ATP-BL) from human leukocytes. DNA Res. 6 (1):29.

[2] Sreedharan et al. (1995) Structure, expression, and chromosomal localization of the type I human vasoactive intestinal peptide receptor gene. Proc. Natl. Acad. Sci. U S A. 92 (7):2939.

[3] Yue et al. (1997) Apoptosis and necrosis in the newborn piglet brain following transient cerebral hypoxia-ischaemia. Neuropathol. Appl. Neurobiol. 23 (1):16.

[4] Zammetto et al. (2005) Unambiguous identification of expressed MAGE-A genes on a DNA microarry. Clin Chem 51:2420.

[5] Grimmond et al. (2001) Expression Profiling with DNA Microarray's: A User's Perspective and Guide. In: DNA Microarrays: Gene Expression Applications (Jordan). Springer Verlag Berlin Heidelberg.