Fluorescein-12-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: C39H41N4O21P3 (free acid)

Peso molecular: 994.69 g/mol (free acid)

Pureza: ≥ 95 % (HPLC)

Forma: solution in water

Concentração: 1.0 mM - 1.1 mM

pH: 7.5 ±0.5

Propriedades espectroscópicas: λexc 492 nm, λem 517 nm, ε 83.0 L mmol-1 cm-1 (Tris-HCl pH 7.5)

Formulários:
Incorporation into DNA/cDNA by
- PCR with Taq polymerase ^{in-house data, [1]}
- Nick Translation with DNAse I/ DNA Polymerase I ^{in-house data}

Descrição:
Fluorescein-12-dUTP is recommended for direct enzymatic labeling of DNA/cDNA e.g. by PCR and Nick Translation. It is incorporated as substitute for its natural counterpart dTTP. The resulting Dye-labeled DNA/cDNA probes are ideally suited for fluorescence hybridization applications such as FISH or microarray-based gene expression profiling.Optimal substrate properties and thus labeling efficiency is ensured by an optimized linker attached to the C5 position of uridine.
Recommended Fluorescein-12-dUTP/dTTP ratio for PCR and Nick Translation: 30-50% Fluorescein-12-dUTP/ 70-50% dTTP

Please note: Protect the Dye-labeled dUTP from exposure to light and carry out experimental procedures in low light conditions. The optimal final concentration of the Dye-labeled dUTP may very depending on the application and assay conditions. For optimal product yields and high incorporation rates an individual optimization of the Dye-labeled-dUTP/dTTP ratio is recommended.

Referências selecionadas:
[1] Wiegant et al. (2001) Probe Labeling and Fluorescence in situ Hybridization. Current Protocols in Cytometry 8:8.3.1.