The Ambion kit is for labeling DNA using a random-priming method, which has technical improvements over existing methods including the use of a high-purity, exonuclease-free Klenow and Random Decamers to produce probes with greater than 109 cpm/μg in 10 min reactions. The kit includes sufficient reagents for 30 reactions.
Don't Trade Specific Activity for Yield
There is a trade-off between probe yield and probe specific activity when using the random-priming method for labeling DNA. Both limiting nucleotide and template mass affect probe yield. Until the labeled nucleotide becomes limiting, the larger the amount of DNA template used, the greater the yield of probe. However, once the labeled nucleotide becomes limiting, additional template will only result in lower specific activity, since the unlabeled template competes with the labeled probe for target.
Maximal Yields of High Specific Activity Probes
The DECAprime™ II DNA Labeling Kit produces probes with maximum specific activity even when the DNA template is impure or the quantity is unknown or very low. Data shows that low template amounts require long incubation times to reach maximum specific activity. Under these conditions, extended reaction times will increase both the yield and the specific activity of the probe.
The NucAway™ Spin Columns (Cat. No. AM10070) are recommended for probe purification.
- Fast, 10 min reaction time
- Elimination of Klenow exonuclease activity maximizes specific activity
- Improved labeling kinetics maximizes yields
- Flexible: both -dATP and -dCTP buffers supplied with each kit
- Decamers produce greater primer-template stability
DNA and RNA Purification and Analysis, DNA Labeling, Northern Blotting, Nucleic Acid Gel Electrophoresis and Blotting, Nucleic Acid Labeling and Oligo Synthesis, Southern Blotting
Shipping Condition: Dry Ice
|Kit, DNA Labeling|
|DNA and RNA Purification and Analysis, DNA Labeling, Northern Blotting, Nucleic Acid Gel Electrophoresis and Blotting, Nucleic Acid Labeling and Oligo Synthesis, Southern Blotting|
|Exo– Klenow, 10X Decamer Solution, 5X Reaction Buffer minus dATP, 5X Reaction Buffer minus dCTP, DECAtemplate GAPDH-M, and 0.5 M EDTA, Nuclease-free water|