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Biotin DecaLabel™ DNA Labeling Kit

#K0651	for 10 reactions
#K0652	for 30 reactions
Related Documents (in pdf, 30-50 KB): Detailed Protocol: #K0651, #K0652 MSDS (English) MSDS (English-USA) MSDS (German) Flyer

 

Features

• 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.

Biotin DecaLabel™ DNA Labeling Kit, flyer in pdf, 74 KB

Description
The 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 originally developed by Feinberg and Vogelstein (1, 2). The primary improvement over traditional random-prime kits involves the use of random decamers instead of hexamers to ensure more efficient annealing with DNA at 37°C. Klenow Fragment, exo- is also included in the kit; this genetically engineered enzyme has no detectable exonuclease activity. Therefore, the enzyme does not degrade the labeled probe during reaction, which results in a high labeling yield even with low amounts of template. As a result, DNA fragments of any length can be uniformly labeled. Biotin-labeled DNA is detected with the Biotin Chromogenic Detection Kit or with conventional biotin-avidin or biotin-streptavidin detection systems.

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 (see Fig.1 below).

Applications
Generation of biotin-labeled DNA probes for a variety of non-radioactive hybridization experiments, including Southern and Northern blots, colony hybridizations, dot/slot blots and in situ hybridizations.

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 (0.1-0.03 pg) are detected after 16 hours of color development with Biotin Chromogenic Detection Kit.

Components of the Kit

• Klenow Fragment, exo-
• Decanucleotide in Reaction Buffer
• Biotin Labeling Mix
• Control Template
• Biotin-labeled DNA
• Water, nuclease-free
• Detailed Protocol (in pdf, ~25 KB):
  • #K0651
  • #K0652

Figure 1. DNA labeling by random primer method.
* [alfa-³²P]-dNTP, [alfa-³²P]-dNTP or biotin-dUTP can be used.

Figure 2. Dot-blot hybridization with a biotin-labeled probe prepared with  the Biotin DecaLabel™ DNA Labeling Kit.
Biotin-labeled lambda DNA/HindIII was 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.

Related Products

Biotin Chromogenic Detection Kit SensiBlot™ Plus Nylon Membrane BCIP-T NBT Genomic DNA Purification Kit DNA Gel Extraction Kit Agarase

References

1. Feinberg, A.P., Vogelstein, B., A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity, Biochem., 132, 6-13, 1983.
2. Feinberg, A.P., Vogelstein, B., A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity, Addendum, Biochem., 137, 266-267, 1984.

Updated balandžio 24, 2008 18:51