Biotin DecaLabel™ DNA Labeling Kit

Detailní popis

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

• Klenow Fragment, exo-
• Decanucleotides in Reaction Buffer
• Biotin Labeling Mix
• Control Template
• Biotin-labeled DNA
• Biotin Labeling Mix
• Detailed Protocol

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. [α-³²P]-dNTP, [α-³³P]-dNTP, biotin-dUTP, fluorescein-, aminoallyl- or DIG-dUTP can be used.

CategoryName
Hazardous	No
Hazardous:	No
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.
Shelf Life:
Shipping Condition:
Storage Condition	-20 C
Storage Condition:	-20 C