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The European Physical Journal D

, Volume 60, Issue 1, pp 31–36 | Cite as

Preparation of DNA films for studies under vacuum conditions

The influence of cations in buffer solutions
  • M. A. ŚmiałekEmail author
  • R. Balog
  • N. C. Jones
  • D. Field
  • N. J. Mason
Topical issue on Molecular level assessments of radiation biodamage

Abstract

Experiments were carried out to determine the optimum conditions required for the preparation of uniform films of supercoiled plasmid DNA to be used in irradiation experiments under high vacuum conditions. Investigations reveal that significant damage to the DNA molecules occurs due to the evacuation process when films were formed from DNA samples in ultra high purity water only. A variety of bases were tested for their possible protective capabilities and sodium hydroxide solution was found to be the most effective in maintaining the supercoiled structure of plasmid DNA during the preparation process. Using a transmission electron microscope we also examined the structure of the DNA films which are formed upon evacuation and how the proposed adducts influence the preparation process. It was found that the addition of bases cause the DNA to aggregate, noting that a base is required for the stability of the DNA molecules. The experimental results presented in this paper show that it may not be possible to perform experiments on so-called pure DNA under vacuum with no stabilizers being added to the sample before the evacuation process.

Keywords

Tantalum Evacuation Process High Vacuum Condition Trizma Base Tantalum Foil 
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.

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References

  1. 1.
    B. Boudaïffa, P. Cloutier, D. Hunting, M.A. Huels, L. Sanche, Science 287, 1658 (2000) ADSCrossRefGoogle Scholar
  2. 2.
    C.A. Hunniford, R.W. McCullough, R.J.H. Davies, D.J. Timson, Biochem. Soc. Trans. 37, 893896 (2009) CrossRefGoogle Scholar
  3. 3.
    M. Folkard, K.M. Prise, B. Brocklehurst, B.D. Michael, J. Phys. B At. Mol. Opt. Phys. 32, 27532761 (1999) CrossRefGoogle Scholar
  4. 4.
    K. Dose, A. Biegerdose, O. Kerz, M. Gill, Orig. Life Evol. Biosph. 21, 177 (1991) ADSCrossRefGoogle Scholar
  5. 5.
    J.A. Wyer, K.T. Butterworth, D.G. Hirst, C.J. Latimer, E.C. Montenegro, M.B. Shah, F.J. Currell, Phys. Med. Biol. 54, 4705 (2009) CrossRefGoogle Scholar
  6. 6.
    M. Folkard, K.M. Prise, B. Vojnovic, S. Davies, M.J. Roper, B.D. Michael, Int. J. Radiat. Biol 64, 651 (1993) CrossRefGoogle Scholar
  7. 7.
    M. Folkard, K.M, Prise, B. Vojnovic, B. Blocklehurst, B.D. Michael, Int. J. Radiat. Biol 76, 763 (2000) CrossRefGoogle Scholar
  8. 8.
    M.A. Huels, B. Boudaiffa, P. Cloutier, D. Hunting, L. Sanche, J. Am. Chem. Soc. 125, 4467 (2003) CrossRefGoogle Scholar
  9. 9.
    S. Even-Chen, Y. Barenholz, Biochim. Biophys. Acta 1509, 176 (2000) CrossRefGoogle Scholar
  10. 10.
    A. Wu, Z. Li, L. Yu, H. Wang, E. Wang, Anal. Sci. 17, 583 (2001) CrossRefGoogle Scholar

Copyright information

© EDP Sciences, SIF, Springer-Verlag Berlin Heidelberg 2010

Authors and Affiliations

  • M. A. Śmiałek
    • 1
    Email author
  • R. Balog
    • 2
  • N. C. Jones
    • 3
  • D. Field
    • 2
  • N. J. Mason
    • 4
  1. 1.Atomic Physics Division, Department of Atomic Physics and LuminescenceFaculty of Applied Physics and Mathematics, Gdańsk University of TechnologyGdańskPoland
  2. 2.Department of Physics and AstronomyAarhus UniversityAarhus CDenmark
  3. 3.Institute for Storage Ring Facilities, Aarhus UniversityAarhus CDenmark
  4. 4.Department of Physics and AstronomyThe Open UniversityMilton KeynesUK

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