Advertisement

A Simple and Rapid Method for Mini Preparation of High Molecular Weight DNA from Certain Acarines, Bacteria and Soybean

  • Z. A. El-FikyEmail author
Research Article

Abstract

A simple, rapid and low-cost method for the isolation of high molecular weight DNA from small amounts of cells or tissue is described. The method does not require liquid nitrogen, proteinase K or costly buffers. This method was successfully applied in the isolation and purification of DNA from eggs of Hyalomma excavatum, H. dromedarii, Argas persicus, A. hermanii and Boophilus annulatus, in addition to Escherichia coli, actinomycetes (Nocardia sp.) and leaves of soybean (Glycine max L.). The DNA concentration was calculated from the optical density (OD) value at wavelength 260 nm and the ratios OD260/OD280 and OD260/OD230 determined. The quality and amount of DNA obtained by this method were comparable to those of DNA isolated from equal amounts of material using a commercial genomic DNA purification kit.

Key Words

DNA extraction simple method Hyalomma excavatum H. dromedarii Argas persicus A. hermanii Boophilus annulatus Escherichia coli Nocardia sp Glycine max 

Résumé

Cette publication présente une méthode simple, rapide et peu coûteuse d’isolement de l’ADN de poids moléculaire élevé à partir d’une petite quantité de cellules ou de tissus. La méthode ne nécessite ni hydrogène liquide, ni protéinase K, ni même de tampons coûteux. Cette méthode a été appliquée avec succès pour l’isolement et la purification d’ADN d’oeufs de Hyalomma excavatum, de H. dromedarii, d’Argas persicus, d’A. hermanii et de Boophilus annulatus, ainsi que d’Escherichia coli, de l’actinomycète (Nocardia sp.) et de feuilles de soja (Glycine max L.). La concentration d’ADN est calculée par la méthode des densités optiques (DO) pour une longueur d’onde de 260 nm et les rapports DO260/DO280 et DO260/DO230 sont déterminés. La qualité et la quantité d’ADN obtenues par cette méthode sont comparables à celles isolées de quantités identiques de matériel à partir d’un kit commercial de purification d’ADN génomique.

Mots Clés

extraction d’ADN méthode simple Hyalomma excavatum H. dromedarii Argas persicus A. hermanii Boophilus annulatus Escherichia coli Nocardia sp Glycine max 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Aboytes-Torres R. (1992) Development and practical applications of a PCR based diagnostic system for the study of chronic bovine anaplasmosis. PhD Dissertation. University of Missouri, Columbia, MO, USA.Google Scholar
  2. Bloom R.A., Mullin B.C. and Tate R.L. (1989) DNA restriction patterns and solution hybridization studies of Frankia isolates from Myrica pensylvanica (bayberry). Appl. Environ. Microbiol. 55, 2155–2160.PubMedPubMedCentralGoogle Scholar
  3. Couch J.A. and Fritz P.J. (1990) Isolation of DNA from plants high in polyphenolics. Plant Mol. Biol. Rep. 8, 8–12.CrossRefGoogle Scholar
  4. de Kok J.B., d’Oliveira C. and Jongejan F. (1993) Detection of the protozoan parasite Theileria annulata in Hyalomma ticks by the polymerase chain reaction. Exp. Appl. Acarol. 17, 839–846.CrossRefGoogle Scholar
  5. Dellaporta S.L., Wood J. and Hicks J.B. (1983) A plant DNA mini preparation. Version III, Plant Mol. Biol. 1, 19–21.CrossRefGoogle Scholar
  6. d’Oliveira C., van der Weide M., Jacquiet P. and Jongejan F. (1997) Detection of Theileria annulata by PCR in ticks collected from cattle in Mauritania. Exp. Appl. Acarol. 21, 279–291.CrossRefGoogle Scholar
  7. Gage K.L., Schrumpf M.E., Karstens R.H., Burgdorfer W. and Schwan T.G. (1994) DNA typing of rickettsiae in naturally infected ticks using a PCR/RFLP system. Amer. J. Trop. Med. Hyg. 50, 247–260.CrossRefGoogle Scholar
  8. Guillemaut P. and Marechal-Drouard L. (1992) Isolation of plant DNA: A fast, inexpensive and reliable method. Plant Mol. Biol. Rep. 10, 60–65.CrossRefGoogle Scholar
  9. Guttman D.S., Wang P.W., Wang I.N., Bosler E.M., Luft B.J. and Dykhuizen D.E. (1996) Multiple infections of Ixodes scapularis ticks by Borrelia burgdorferi as revealed by single strand conformation polymorphism analysis. J. Clin. Microbiol. 34, 652–656.PubMedPubMedCentralGoogle Scholar
  10. Hubbard M.J., Cann K.J. and Wright D.J.M. (1995) Validation and rapid extraction of nucleic acids from alcohol-preserved ticks. Exp. Appl. Acarol. 19, 473–478.PubMedGoogle Scholar
  11. Jhingan A. K. (1994) DNA isolation from animal cells. PATN Patent Bibliographic Information, GenBank No. 05352777.Google Scholar
  12. Jongejan F., van Vliet A.H.M., van der Weide M. and d’Oliveira C. (1998) Detection of two tick-borne pathogens (Theileria annulata and Cowdria ruminantium) by the polymerase chain reaction, pp. 521–525. In Proceedings of the Second International Conference on Tick-borne Pathogens. Kruger National Park. Republic of South Africa, 28 August-1 September 1995.Google Scholar
  13. Junghans H. and Metzlaff M. (1990) A simple and rapid method for the preparation of total plant DNA. Biotechniques 8, 176.PubMedGoogle Scholar
  14. Kamalay J.C., Tejwani R. and Rufener G.K. (1990) Isolation and analysis of genomic DNA from single seeds. Crop Scienc. 30, 1079–1084.CrossRefGoogle Scholar
  15. Katterman F.R.H. and Shattuck V.I. (1983) An effective method of DNA isolation from the mature leaves of Gossypium species that contain large amounts of phenolic terpenoids and tannins. Preparative Biochemistr. 13, 347–359.CrossRefGoogle Scholar
  16. Marmur J. (1961) A procedure for the isolation of deoxyribonucleic acid from microorganisms. J. Mol. Biol. 3, 208–218.CrossRefGoogle Scholar
  17. Mauel M.J., Carlton S.J. and Mather T. N. (1999) Polymerase chain reaction detection efficiency of the human granulocytic ehrlichiosis agent (Rickettsiaceae: Ehrlichieae) in ticks (Acari: Ixodidae) is dependent on the DNA extraction method. J. Med. Entomol. 36, 649–652.CrossRefGoogle Scholar
  18. Peterson D.G., Boehm K.S. and Stack S.M. (1997) Isolation of milligram quantities of nuclear DNA from tomato (Lycopersicon esculentum), a plant containing high levels of polyphenolic compounds. Plant Mol. Biol. Rep. 15, 148–153.CrossRefGoogle Scholar
  19. Ramelow C., Suss J., Berndt D., Roggendorf M. and Schreier E. (1993) Detection of tick-borne encephalitis virus RNA in tick (Ixodes ricinus) by the PCR. J. Virol. Meth. 45, 115–119.CrossRefGoogle Scholar
  20. Sambrook J., Fritsh E.F. and Maniatis T. (1989) Molecular Cloning: A Laboratory Manual, 2nd edition (Edited by C. Nolan). Cold Spring Harbor Laboratory Press, USA.Google Scholar
  21. Shatters R.G., Schweder M.E., West S. H., Abdelghany A. and Smith R.L. (1995) Environmentally induced polymorphisms detected by RAPD analysis of soybean seed DNA. Seed Sci. Res. 5, 109–116.CrossRefGoogle Scholar
  22. Sparagano O.A.E., Allsopp M.T.E.P., Mank R.A., Rijpkema S.G.T., Figueroa J.V. and Jongejan F. (1999) Molecular detection of pathogen DNA in ticks (Acari: Ixodidae): A review. Exp. Appl. Acarol. 23, 929–960.CrossRefGoogle Scholar
  23. Walbot V. (1989) Preparation of DNA from single rice seedlings. Rice Genetics Newsl. 5, 149–151.Google Scholar
  24. Ward V.K., Marriott A.C., Booth T.F., El-Ghorr A.A. and Nuttall P.A. (1990) Detection of an arbovirus in an invertebrate and a vertebrate host using the PCR. J. Virol. Meth. 30, 291–300.CrossRefGoogle Scholar
  25. Watson J.C. and Thompson W.F. (1986) Purification and restriction endonuclease analysis of plant nuclear DNA. Methods Enzymol. 118, 57–79.CrossRefGoogle Scholar
  26. Williams J.G.K., Kubelik A.R., Livak K.J., Rafalski J.A. and Tingey S.V. (1990) DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res. 18, 6531–6535.CrossRefGoogle Scholar
  27. Winfrey M.R., Rott M.A. and Wortman A.T. (1997) Unravelling DNA: Molecular Biology for the Laboratory. Prentice Hall Inc., Simon & Schuster/A. Viacom Company, New Jersey. 396 pp.Google Scholar

Copyright information

© ICIPE 2003

Authors and Affiliations

  1. 1.Genetics Department, Faculty of AgricultureFayoum Branch Cairo UniversityFayoumEgypt

Personalised recommendations