Employing click chemistry, FISH probes are synthesized via PCR including modified nucleotides. In the triphosphate mixtures dTTP is replaced by C8-Alkyne-dUTP or dCTP, respectively. Both mixtures can be used in standard PCR with the optimized Baseclick polymerase.
The resulting PCR products contain alkyne moieties, which react with azido-derivatives of fluorescent dyes, haptens and others in a highly selective fashion under benign click reaction conditions. After hybridization of these alkyne-modified probes to the target sequence, conditions. After hybridization of these alkyne-modified probes to the target sequence, detection is carried out by a post labelling click reaction followed by fluorescence microscopy.
POST-HYBRIDIZATION CLICK KITS /PROBES
The baseclick's Click-FISH kits provide all the reagents required for the generation of highly labeled fluorescence in situ hybridization probes.
The Click-Fish Kit contains chemicals to perform 40 PCR reactions (25µl each) and 500 different independent labeling reactions(FISH experiments).