DecaLabel™ DNA Labeling Kit

Detailní popis

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

• Klenow Fragment, exo-
• Decanucleotides in Reaction Buffer
• Mix A (-dATP)
• Mix C (-dCTP)
• dNTP Mix
• Control Template
• Water, nuclease-free
• Detailed Protocol

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).