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Biotin DecaLabel DNA Labeling Kit


Biotin DecaLabel DNA Labeling Kit is an advanced system for the efficient synthesis of biotin-labeled DNA probes.
  
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The Thermo Scientific Biotin DecaLabel DNA Labeling Kit is an advanced system for the efficient synthesis of biotin-labeled DNA probes, based on an improved random-primed labeling method developed by Feinberg and Vogelstein (see References 1, 2). The primary improvement over traditional random-primed method involves the use of random decamers instead of hexamers, ensuring more efficient annealing with DNA at 37°C. Klenow Fragment, exo-, included in the kit, is genetically engineered enzyme with no detectable exonuclease activity. The enzyme does not degrade the labeled probe during reaction, which results in a high labeling yield even with low amounts of template. You can uniformly label any length DNA fragments.

Biotin-labeled DNA is detected with the Biotin Chromogenic Detection Kit or conventional biotin-avidin or biotin-streptavidin detection systems.

Highlights

  • Non-radioactive labeling of DNA
  • Efficient priming of labeling reactions with random decamers
  • High yields with Klenow Fragment, exo-: no degradation of a labeled probe during reaction

Applications

  • Generation of biotin-labeled DNA probes for a variety of non-radioactive hybridization experiments, including 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 molecule, and new strands are synthesized by the Klenow Fragment, exo- in the presence of biotin-dUTP. During this reaction, the biotinylated nucleotides are incorporated into the newly synthesized complementary DNA strand.

DNA labeling by the random primed method

DNA labeling by the random primed method. [α-32P]-dNTP, [α-33P]-dNTP, biotin-dUTP, fluorescein-, aminoallyl- or DIG-dUTP can be used.

  
Storage Condition-20 C
HazardousNo
Quality Control
  • The kit is tested in a control labeling reaction.
  • The biotin-labeled DNA probe is used in dot-blot.
  • Low amounts of homologous DNA (0.1 to 0.03 pg) are detected after 16 hours of color development with Biotin Chromogenic Detection Kit.
Dot-blot hybridization with a biotin-labeled probe

Dot-blot hybridization with a biotin-labeled probe

Dot-blot hybridization with a biotin-labeled probe

Dot-blot hybridization with a biotin-labeled probe. Lambda DNA/HindIII was biotin-labeled with the Biotin DecaLabel DNA Labeling Kit and used as a hybridization probe in a dot-blot of the homologous DNA on SensiBlot Plus Nylon Membrane. The blot was developed with the Biotin Chromogenic Detection 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).