Fluorescein-12-dCTP

excitation and emission spectrum of 5/6-FAM

For general laboratory use.

Please centrifuge briefly before opening (volume ≤2 ml).

Description:
Fluorescein-12-dCTP 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 dCTP. 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 cytidine.

Recommended Fluorescein-12-dCTP/dCTP ratio for PCR and Nick Translation: 30-50% Fluorescein-12-dCTP-/ 70-50% dCTP

Please note: Protect the Dye-labeled dCTP from exposure to light and carry out experimental procedures in low light conditions. The optimal final concentration of the Dye-labeled dCTP may very depending on the application and assay conditions. For optimal product yields and high incorporation rates an individual optimization of the Dye-labeled-dCTP/dCTP 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: C₃₉H₄₀N₅O₂₀P₃

Molecular Weight: 991.68 g/mol

Exact Mass: 991.15 g/mol

Purity: ≥ 95 % (HPLC)

Form: solution in 10 mM Tris-HCl

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}
- Nick Translation with DNAse I/ DNA Polymerase I ^{in-house data}