For generation of highly sensitive probes for fluorescence in situ hybridization (FISH). The Nick Translation Mix is designed for direct fluorophore-labeling of in situ probes. Fluorescein-12-dUTP and Tetramethyl-Rhodamine-5-dUTP from Roche Applied Science or other commercially available fluorophor-labeled nucleotides can be combined with the Nick Translation Mix. Direct fluorophore-labeled in situ probes are used for the detection of multi copy or very large hybridization targets on metaphase chromsomes or interphase nuclei. For a standard labeling reaction using 1 μg template in 20 μl total reaction volume, 4 μl of 5x concentrated fluorophore labeling mix are required.
一般的な説明
Optimized enzyme mixture: Nick translation utilizes a combination of DNase and DNA Polymerase to nick one strand of the DNA helix, then incorporates labeled nucleotides as the polymerase examines, or "proofreads" the nicked site. Individual templates produce consistent results in the standard 90-minutes reaction, and result in an average probe length of 200 base pairs up to 500 base pairs. Assay Time: 100 minutes
