5-Propargylamino-dCTP-Cy3

excitation and emission spectrum of Cy3

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: C₄₃H₅₅N₆O₂₀P₃S₂ (free acid)

Molecular Weight: 1132.97 g/mol (free acid)

Exact Mass: 1132.21 g/mol (free acid)

Purity: ≥ 95 % (HPLC)

Form: solution in 10 mM Tris-HCl

Color: pink

Concentration: 1.0 mM - 1.1 mM

pH: 7.5 ±0.5

Spectroscopic Properties: λ_exc 550 nm, λ_em 570 nm, ε 150.0 L mmol^-1 cm^-1 (Tris-HCl pH 7.5)

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

Description:
5-Propargylamino-dCTP-Cy3 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 Propargylamino-dCTP-Cy3/dCTP ratio for PCR and Nick Translation: 30-50% Propargylamino-dCTP-Cy3/ 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.

BIOZ Product Citations:

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Selected References:
[1] Walsh et al. (2017) Measurement of incorporation kinetics of non-fluorescent native nucleotides by DNA polymerases using fluorescence microscopy. Nucleic Acids Res. 45 (21):e175.
Ramsay et al. (2010) CyDNA: Synthesis and Replication of Highly Cy-Dye
Substituted DNA by an Evolved Polymerase. J. Am. Chem. Soc. 132 (14):5096.