HighFidelity Digoxigenin PCR Labeling Kit

HighFidelity Digoxigenin PCR Labeling Kit is designed to produce randomly Digoxigenin-modified DNA probes by PCR. Such probes are ideally suited for in situ hybridization and Northern Blot experiments. PCR-based labeling is superior to random-primed labeling by Klenow fragment if template amounts are limited or amplification of a specific DNA fragments is required. The labeling principle is similar to the underlying labeling principles of PCR DIG Labeling Mix (Roche) and PCR DIG Probe Synthesis Kit (Roche) with the exception that an alkali-stable version of DIG-11-dUTP is used (DIG label removal not feasible) . DIG-11-dUTP is efficiently incorporated into DNA as substitute for its natural counterpart dTTP using an optimized reaction buffer and a High Fidelity Polymerase. 35% DIG-11-dUTP substitution typically results in an optimal balance between reaction and labeling efficiency. Individual optimization of DIG-11-dUTP/dTTP ratio however, can easily be achieved with the single nucleotide format. The resulting Digoxigenin-modified DNA probe can subsequently be detected by HRP-or AP-modified Digoxigenin antibody. The kit contains sufficient reagents for 35 labeling reactions (S-Pack) or 175 labeling reactions (L-Pack) of 20 μl each (35% DIG-11-dUTP substitution, 100 μM dATP/dGTP/dCTP, 65 μM dTTP, 35 μM DIG-11-dUTP) .