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DecaLabel DNA Labeling Kit

Store at -20°C

DecaLabel DNA Labeling Kit is a complete system for fast synthesis of radiolabeled DNA for hybridization experiments.
  
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The Thermo Scientific DecaLabel DNA Labeling Kit is an advanced system for fast synthesis of radiolabeled DNA probes of high specific activity. It's based on the improved random-primed method developed by Feinberg and Vogelstein (see References 1, 2).

The primary improvement over traditional random-primer kits involves the use of random decamers instead of hexamers to ensure more efficient annealing with DNA at 37°C. In addition, Klenow Fragment, exo- has been genetically engineered to remove exonuclease activity. The improved enzyme does not degrade the labeled probe during reaction and allows high labeling yields even from low template amounts. DNA fragments of any length can be uniformly labeled. Two labeling mixes are provided for flexibility in using either radiolabeled dATP or dCTP.

Highlights

  • High specific radioactivity of probes – greater than 1 x 109 dpm/µg DNA
  • Fast – 5 minutes at 37°C
  • High yields with Klenow Fragment, exo- no degradation of a labeled probe during reaction
  • Flexible – suitable for either radiolabeled dATP or dCTP

Applications

  • Generation of radiolabeled DNA probes for use in a variety of hybridization experiments: Southern and Northern blots, colony/plaque hybridizations, dot/slot blots, and in situ hybridizations.

Includes

Principle

Random decamers are annealed to a denatured template DNA, and radiolabeled dNTPs are then incorporated into new DNA strands by Klenow Fragment, exo- (see Figure 1 under Supporting Data).

  
Storage Condition-20 C
HazardousNo
Quality ControlThe kit is tested in a control labeling reaction. The typical specific radioactivity of the synthesized probe is > 1 x 109 dpm/µg DNA.
Effect of time, amount of template on the specific radioactivity of a DNA probe

Effect of time, amount of template on the specific radioactivity of a DNA probe

Effect of time, amount of template on the specific radioactivity of a DNA probe

Figure 1. Effect of reaction time and amount of template on the specific radioactivity of a DNA probe.
Varying amounts of Lambda DNA/HindIII fragments were labeled with 50 µCi [α-33P]-dCTP (2500 Ci/mmol) according to the protocol supplied with the kit.


References

  1. A. P. Feinberg, B. Vogelstein, A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Biochem. 132, 6-13 (1983).

  2. A. P. Feinberg, B. Vogelstein, A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity, Addendum. Biochem. 137, 266-267 (1984).