Fluorescein-12-dUTP

Empirical formula C₃₉H₃₈N₄O₂₁P₃
Molar mass (M) 991,66 g/mol
Storage temp.: -20 °C
Transport temp.: cooled

ε₄₉₅ (pH 9) = 70.0 E x mmol^-1 x cm^-1; pH: 7.5 ±0.2

Fluorescein-12-dUTP is able to replace dTTP in growing DNA-strands and is used for efficient non-radioactive DNA-labelling. Detection of labelled nucleic acids can be done by direct analysis of fluorescent signals (e.g. by fluorescence microscopy) or by incubation with AP-coupled anti-fluorescein antibodies.Fluorescein-12-dUTP can also be used for double staining techniques in combination with rhodamine-12-dUTP or biotin-11-dUTP and corresponding antibodies or streptavidin-complexes, respectively.

Excitation: 494 nm
Emission: 520 nm

Applications:

Non-radioactive labelling of DNA by enzymatic reactions, e.g. PCR, reverse transcription, nick-end-translation, end-labelling or random-primed DNA-labelling. Incorporation can be done with all established DNA-polymerases (e.g. Taq-polymerase, T4 DNA-polymerase, Klenow fragment).

Testet for the lack of endo-, exodeoxyribonuclease, ribonuclease and phosphatase.

Figure:

Detection of chromosome wcp21q (whole chromosome paint, red arrows) and band 11q23 (BAC, bacterial artificial chromosome, white arrows) by FISH, labelling with fluorescein-12-dUTP.

_{With kind permission of PD Dr. Thomas Liehr, Institute of Human Genetics and Anthropology, Friedrich-Schiller University, Jena.}