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DNA probes up to 4 kbp: Efficient Cy®-, ATTO™- or Alexa Fluor®-dye labeling

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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.

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We now offer HighFidelity PCR Labeling Kits...

  • ...for both fluorescent (e.g Cy3 or Alexa Fluor 488), Biotin and Digoxigenin labeling
  • ...suitable for preparation of both short (>100 bp) and long DNA probes (up to 4 kbp tested) due to a High Fidelity Polymerase blend consisting of Taq polymerase and a proofreading enzyme (Fig. 1).
  • ...with convenient configurations for application-specific assay optimization:
    • single nucleotide format for optimization of labeled dUTP/dTTP ratio
    • positive control reaction (amplification of 500 bp fragment)
    • testkits containing selected labeled dUTPs to find the optimal label

Figure 1: High Fidelity Polymerase allows efficient production of 4kbp labeled DNA probes.

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.

Table 1: Overview on available HighFidelity PCR Labeling Kits (* AF: Alexa Fluor)

Dr. Barbara Zschoernig

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