Nick translation is the method of choice for the generation of labeled DNA probes (100- 500 bp) from large templates such as genomic DNA, cosmids, BAC clones. It is based on the concerted activities of DNAse I (introduction of random single-strand breaks (“nicks”) into double-stranded DNA) and Polymerase I that incorporates labeled dUTPs instead of their natural counterpart dTTP at the “nicked” sites thus creating ideal in situ hybridization probes. There is no net DNA synthesis in contrast to PCR-based labeling approaches.
The combination of (poly)-HRP-conjugated Biotin/Digoxigenin detection reagents with AF 488 (Alexa Fluor® 488)-, AF546 (Alexa Fluor® 546)- or AF594 (Alexa Fluor® 594)-labeled tyramide further increases detection sensitivity of up to 100-fold.
Fluorescent Nick Translation Labeling Kits are available as well.