Digoxigenin NT Labeling Kit

Digoxigenin Nick Translation Labeling Kit contains all reagents (except template and materials for purification of the probe) required for nick translation-based Digoxigenin labeling of DNA providing a highly efficient, easy-to-perform and rapid labeling technology. The labeling principle is similar to the DIG-Nick Translation Mix (Roche). Nick translation labeling is based on the reverse activities of Polymerase I and DNase I. DNase I is able to introduce randomly distributed nicks to double stranded DNA at low enzyme concen-trations. The 5'→3' exonuclease activitiy of Polymerase I removes nucleotides from the 3' side of the nick while synthesizing a partial new complementary strand using the 3'-OH termini as primer. In the presence of labeled dUTP Polymerase I incorporates labeled dUTP instead of dTTP. The DIGX NT labeling mix contains an optimized Digoxigenin-labeled dUTP for incorporation into DNA by nick translation using DNA Polymerase I. Digoxigenin-11-dUTP is enzymatically incorporated into DNA as sub-stitute for its natural counterpart dTTP. Optimal substrate properties and thus labeling efficiency as well as an efficient detection of the Digoxigenin moiety is ensured by an 11-atom linker attached to the C5 position of uridine. The well balanced polymerase/nuclease activities of the en-zyme mix ensure the generation of highly labeled double stranded DNA fragments. The resultant Biotin-labeled DNA is suitable for application in FISH and other nucleic acid hybridization assays.