Biotin-11-dUTP

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: C28H45N6O17P3S (free acid)

Peso molecular: 862.67 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 240/289 nm, ε 10.7/7.1 L mmol-1 cm-1 (Tris-HCl pH 7.5)

Formulários:
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 3'-5' exo^{- [5]}
- 3’-End Labeling with Terminal deoxynucleotidyl Transferase (TdT)^[6]
- Reverse Transcription with MMLV Reverse Transcriptase ^[7]

Descrição:
Biotin-11-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 11-atom linker attached to the C5 position of uridine.

Recommended Biotin-11-dUTP/dTTP ratio for PCR and Nick Translation: 50% Biotin-11-dUTP/ 50% dTTP

Please note: The optimal final concentration of Biotin-11-dUTP may very depending on the application and assay conditions. For optimal product yields and high incorporation rates an individual optimization of the Biotin-11-dUTP/dTTP ratio is recommended.

Produtos relacionados:

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

Referências selecionadas:
[1] Day et al. (1990) Synthesis in vitro and application of biotinylated DNA probes for human papilloma virus type 16 by utilizing the polymerase chain reaction. Biochem J 267:119.

[2] Ried et al. (1992) Simultaneous visualization of severe different DNA probes by in situ hybridization using combinatorial fluorescence and digital imaging microscopy. Proc. Natl. Acad. Sci. USA 89:1388.

[3] Langer et al. (1981) Enzymatic synthesis of biotin-labeled polynucleotides: novel nucleic acid affinity probes. Proc Natl Acad Sci USA 78:6633.

[4] Brigati et al. (1983) Detection of viral genomes in cultured cells and para nembedded tissue sections using biotin-labled hybridization probes. Virology 126:32.

[5] Day et al. (2008) Microfluidic-based enzymatic on-chip labeling of miRNAs. N Biotechnol 25:142.

[6] Flickinger et al. (1992) Differential incorporation of biotinylated nucleotides by terminal deoxynucleotidyl transferase. Nucleic Acids Res 20:2382.

[7] Anderson et al. (2005) Incorporation of reporter-labeled nucleotides by DNA polymerases. Biotechniques 38:257.

Moritz et al. (2014) Simple methods for the 3' biotinylation of RNA. RNA. 20 (3):421.