Biochemical Genetics

, Volume 31, Issue 7–8, pp 321–328 | Cite as

DNA isolation by a rapid method from human blood samples: Effects of MgCl2, EDTA, storage time, and temperature on DNA yield and quality

  • Debomoy K. Lahiri
  • Bill Schnabel


The isolation of DNA from whole blood by a modified rapid method (RM) was tested using various detergents and buffer conditions. Extraction of DNA with either NP-40 or Triton X-100 gave a high yield of undegraded DNA in less than an hour. The concentration of magnesium ion in the buffers was critical to obtaining intact, high molecular weight (HMW) DNA. Greater than 10 mM MgCl2 led to degradation. Addition of EDTA to the buffer inhibits this degradation. Preparation of DNA from blood stored at room temperature or incubated at 37°C for 24 hr resulted in the same amount and quality of DNA as from samples frozen at −70°C. DNA from blood samples that had undergone more than four freeze-thaw cycles was found to be partially degraded. The modified RM can be applied to extract DNA from as little as 10 µl of blood (340 ng of DNA) and from dried blood samples. DNA samples remained intact and undegraded for longer times when DNA was dissolved in higher concentrations of EDTA.

Key words

high molecular weight DNA MgCl2 integrity of DNA rapid method DNA banking 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Anderson, W. F. (1992). Human gene therapy.Science 256808.Google Scholar
  2. Douglas, A. M., Georgalis, A. M., Benton, L. R., Canavan, K. L., and Atchi, B. A. (1992). Purification of human leucocyte DNA: Proteinase K is not necessary.Anal. Biochem. 201362.Google Scholar
  3. Lahiri, D. K., and Nurnberger, J. I., Jr. (1991). A rapid non enzymatic method for the preparation of HMW DNA from blood for RFLP studies.Nucleic Acids Res. 195444.Google Scholar
  4. Lahiri, D. K., Bye, S., Nurnberger, J. I., Jr., Hodes, M. E., and Crisp, M. (1992). A non-organic and non-enzymatic extraction method gives higher yields of genomic DNA from whole blood samples than do nine other methods tested.J. Biochem. Biophys. Meth. 25193.Google Scholar
  5. McCabe, E. R. B., Huang, S.-Z., Seltzer, W. K., and Law, M. L. (1987). DNA microextraction from dried blood spots on filter paper blotters: potential applications to newborn screening.Hum. Genet. 75213.Google Scholar
  6. Madisen, L., Hoar, D. I., Holroyd, C. D., Crisp, M., and Hodes, M. E. (1987). DNA Banking: The effects of storage of blood and isolated DNA on the integrity of DNA.Am. J. Med. Genet. 27379.Google Scholar
  7. Maniatis, T., Fritsch, E. F., and Sambrook, J. (1982).Molecular Cloning: A Laboratory Manual Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.Google Scholar
  8. Miller, A. D. (1992). Human gene therapy comes of age.Nature 357455.Google Scholar
  9. Miller, S. A., Dykes, D. D., and Polesky, H. F. (1988). A simple salting out procedure for extracting DNA from human nucleated cells.Nucleic Acids Res. 161215.Google Scholar
  10. Saiki, R. K., Bugawan, T. L., Horn, G. T., Mullis, K. B. and Erlich, H. A. (1986). Analysis of enzymatically amplified β-globin and HLA-DQα DNA with allele-specific oligonucleotide probes.Nature 324163.Google Scholar

Copyright information

© Plenum Publishing Corporation 1993

Authors and Affiliations

  • Debomoy K. Lahiri
    • 1
  • Bill Schnabel
    • 1
  1. 1.Laboratory of Molecular Neurogenetics, Institute of Psychiatric Research, Department of PsychiatryIndiana University School of MedicineIndianapolis

Personalised recommendations