HighFidelity Fluorescein PCR Labeling Kit

HighFidelity Fluorescein PCR Labeling Kit is designed to produce randomly Fluorescein-modified DNA probes by PCR. Such probes are ideally suited for Fluorescence in situ hybridization (FISH) and Northern Blot experiments. PCR-based labeling is superior to random-primed labeling with Klenow fragment if template amounts are limited or amplification of a specific DNA fragments is required. Amplification of probes up to 4 kbp is feasible. Fluorescein-12-dUTP is efficiently incorporated into DNA as substitute for its natural counterpart dTTP using an optimized reaction buffer and a High Fidelity Polymerase blend consisting of Taqpolymerase and a proofreading enzyme. 50% Fluorescein-12-dUTP substitution typically results in an optimal balance between reaction and labeling efficiency. Individual optimization of Fluorescein-12-dUTP/dTTP ra-tio however, can easily be achieved with the single nucleotide format. The kit contains sufficient reagents for 10 labeling reactions (S-Pack) or 50 labeling reactions (L-Pack) of 20 µl each (50% Fluorescein-12-dUTP substitution, 100 µM dATP/dGTP/dCTP, 50 µM dTTP, 50 µM Fluorescein-12-dUTP).