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Introduction to Radioactive LabelingRandom Prime (1, 2) is a quick and reliable method for labeling very small quantities of DNA to very high specific activities. It uses labeled nucleotides more efficiently than nick translation and requires no prior purification of DNA fragments separated from low-melting-point agarose. The DNA to be labeled is first denatured and then mixed with oligonucleotides of random sequence in the presence of labeled nucleotide(s) and a DNA polymerase.
These “random oligos” anneal to complementary random sites on the DNA and serve as primers for DNA synthesis during which the label becomes incorporated.
References
- Feinberg, A. P. and Vogelstein, B., Anal. Biochem. 132, 6 (1983).
- Feinberg, A. P. and Vogelstein, B., Anal. Biochem. 137, 266 (1984).
| SELECTION GUIDE - Radioactive Labeling Systems |
| Labeling system | Technology | Nucleotide | Amount of template | Labeling time | Probe specific activity (dpm/µg) | Recommended application |
| Rediprime™ II | Random-prime | dCTP only | 25 ng | 10 min | 2 × 109 | membrane hybridization |
Ready-To-Go™
DNA labeling beads | Random-prime | dCTP only | 10 ng to 1 µg | 5 min | 2 × 109 | membrane hybridization |
| Megaprime™ | Random-prime | any dNTP | 25 ng | 10 min | 2 × 109 | membrane hybridization |
| Nick translation | Nick translation | any dNTP | 1 µg | 2-3 h | 2 × 109 | Production of large
amounts of probe |
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