Labeled DNA probes e.g. for (Fluorescence) in situ hybridization ((F)ISH) are routinely prepared via PCR if template amounts are limited or labeling of sequence-specific DNA fragment is required.
Figure 1: High Fidelity Polymerase allows efficient production of 4 kbp labeled DNA probes.
Taq polymerase- and High Fidelity Polymerase-mediated production of 4 kbp labeled DNA probes has been compared in the presence of varying amounts of labeled dUTP. Visualization of analog incorporation was performed by agarose gel electrophoresis without DNA intercalator. Product formation was visualized by agarose gel electrophoresis with DNA intercalator EtBr. A) Production of 4 kbp ATTO488-labeled DNA probe with dUTP-XX-ATTO-488. B) Production of 4 kbp Digoxigenin-labeled DNA probe with DIG-11-dUTP. Band shift indicates Dig-11-dUTP incorporation. M: DNA Ladder (Cat. No. M-216), 0 - 100 %: Increasing amount of dTTP substitution by labeled dUTP.
Nick Translation Labeling Kits for fluorescent or Hapten Labeling are available as well.
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