Fluorescein-12-dUTP

Excitation and Emission spectrum of 5/6-FAM

For general laboratory use.

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: C₃₉H₄₁N₄O₂₁P₃ (free acid)

Molecular Weight: 994.69 g/mol (free acid)

Exact Mass: 994.15 g/mol (free acid)

Purity: ≥ 95 % (HPLC)

Form: solution in water

Color: yellow-orange

Concentration: 1.0 mM - 1.1 mM

pH: 7.5 ±0.5

Spectroscopic Properties: λ_exc 492 nm, λ_em 517 nm, ε 83.0 L mmol^-1 cm^-1 (Tris-HCl pH 7.5)

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

Description:
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.

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