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Nonradioactive Labeling of DNA

  • Thomas P. McCreery
  • Terrence R. Barrette
Protocol
  • 1k Downloads
Part of the Springer Protocols Handbooks book series (SPH)

Abstract

Many important advances in molecular biology would not have been possible wlthout the use of radioisotopes It is relatively simple to substitute a radtoacttve isotope into a nucleotide to produce a molecule that has the same biological properties as the unlabeled molecule (see Chapter 6) These molecules are mcorporated into DNA sequences by a variety of protocols Unfortunately, with the high efficiency and easy incorporation of radionuchdes comes a Pandora’s Box of difficulties The short half lives of the most commonly used nucleotides (32P,33P, 35S) necessitate that the material be freshly labeled for optimal efficiency This alone makes it difficult for scientists m developing nations to use isotopic techniques. The ability of radiation to penetrate human tissue and cause damage requires that all work be done from behind shielding material Personnel exposure to radiation must be monitored on a regular basis A problem that is now looming large for the use of radiation in the molecular-biology laboratory is the lack of disposal sites for radioactive waste. Many regulatory agencies will not issue licenses to work with radlatton if no disposal site is available

Keywords

Radioactive Waste Disposal Site Intestinal Alkaline Phosphatase Calf Intestinal Alkaline Phosphatase Alkaline Phosphatase Enzyme 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

References

  1. 1.
    McCreery, T and HelentJaris, T (1994) Production of hybrldlzatlon probes by the PCR utlllzing dlgoxlgenm-modified nucleotldes, in Methods zn Molecular Bzology, vol 28 Protocols for Nuclezc Aczd Analyszs by Nonradzoactzve Probes (Isaac, P, ed ), Humana, Totowa, NJ, pp 67–71Google Scholar
  2. 2.
    McCreery, T and HelentJaris, T (1994) Production of hybrldlzatlon probe with dlgoxlgenm-modified nucleotldes by random hexanucleotlde priming, in Methods in Molecular Bzology, vol 28 Protocols for Nucleic Aczd Analyszs by Nonradzoactzve Probes (Isaac, P, ed ), Humana, Totowa, NJ, pp 73–76Google Scholar
  3. 3.
    Karp, A (1994) Labeling of double-stranded DNA probes with blotm, in Methods in Molecular Bzology, vol 28 Protocols for Nuclezc Aczd Analyszs by Nonradzoactzve Probes (Isaac, P, ed ), Humana, Totowa, NJ, pp 83–87Google Scholar
  4. 4.
    Durrant, I and Stone, T (1994) Preparation of horseradish peroxldase-labeled probes, in Methods in Molecular Bzology, vol 28 Protocols for Nucleic Aczd Analyszs by Nonradzoactzve Probes (Isaac, P, ed ), Humana, Totowa, NJ, pp 89–92Google Scholar
  5. 5.
    The Guzde to Non-Radzoactzve Products (1993) Promega Corporation, Madison WI, pp 2–2–2–10Google Scholar
  6. 6.
    McCreery, T and HelentJans, T (1994) Hybrldlzatlon of dlgoxlgenm-labeled probes to Southern blots and detectton by chemdummescence, in Methods in Molecular Bzology, vol 28 Protocols for Nucleic Aczd Analysis by Nonradzoactzve Probes (Isaac, P, ed ), Humana, Totowa, NJ, pp 107–112Google Scholar
  7. 7.
    Durrant, I and Stone, T (1994) Hybrldlzatlon of horseradish peroxldase-labeled probes and detection by enhanced chemllummescence, in Methods in Molecular Bzology, vol 28 Protocols for Nucleic Aczd Analyszs by Nonradzoactzve Probes (Isaac, P, ed ), Humana, Totowa, NJ, pp 127–133Google Scholar
  8. 8.
    McQuald, S, McMahon, J, and Allan, G (1995) A comparison of dlgoxlgemn and blotm labelled DNA and RNA probes for in situ hybrldlzatlon Biotech Histochem 70, 147–154CrossRefGoogle Scholar

Copyright information

© Humana Press Inc , Totowa, NJ 1998

Authors and Affiliations

  • Thomas P. McCreery
    • 1
  • Terrence R. Barrette
    • 1
  1. 1.ImaRx Pharmaceutzcals,IncAZ

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