DIG-Nick Translation Mix has been used for the generation of highly sensitive probes for in situ hybridization labeled with Digoxigenin-11-dUTP. Note: The mix can also be used for filter hybridization techniques, however, for highly sensitive filter hybridization probes, we recommend that you use DIG-High Prime from Roche Applied Science. For nonradioactive labeling of in situ probes with other haptens and fluorophores, Roche Applied Science offers the Biotin-Nick Translation Mix and the Nick Translation Mix (without nucleotides).
일반 설명
DIG-Nick Translation Mix is a convenient enzyme and nucleotide mixture for nick translation. It 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. Large plasmids, cosmids and polymerase chain reaction (PCR) fragments are all regularly used with this method and the DNA can be linearized or supercoiled. Individual templates produce consistent results in the standard 90-minutes reaction and result in an average probe length of 200 bp up to 500 bp. The molar ratio of digoxigenin (DIG)-11-deoxyuridine triphosphate (dUTP) to deoxythymidine triphosphate (dTTP) is adjusted to ensure that every 20th to 25th nucleotide in the newly synthesized DNA is modified with DIG. This density of haptens in the DNA yields the highest sensitivity in the immunological detection reaction.
