Summary
We describe methods for extracting genomic DNA from a small amount of whole blood or cultured amniocytes. Nuclear DNA was extracted from whole blood spotted on blotting paper. Relatively large molecules of DNA with the average amount of 7–9 μg was extracted from 1 ml of blood spotted and stored for at most two years, being roughly 1/3 of that extracted directly from fresh whole blood. The estimated minimum amount of whole blood that gives a suitable autoradiogram of Southern hybridization was 0.3 ml. Another series of amounts of whole blood or an amniocyte suspension were molded in low-melting agarose into an 100 μl gel block. The DNA extracted from a block that was made from at least 0.25 ml of whole blood, or from 1.25×105 amniocytes (equivalent to 1/8 of the number of confluent cells in a 25 cm2 culture flask) resulted in one suitable Southern analysis. Both methods described here are applicable to the diagnosis of newborns and/or fetuses at risk of a genetic disease and to the diagnosis of a patient from whom a large amount of blood material is difficult to obtain. These methods also make a long-way transportation of the materials possible.
Similar content being viewed by others
Article PDF
References
Cooper, D.N. and Schmidtke, J. 1986. Diagnosis of genetic disease using recombinant DNA.Hum. Genet. 73: 1–11.
Cooper, D.N. and Schmidtke, J. 1987. Diagnosis of genetic disease using recombinant DNA. Supplement.Hum. Genet. 77: 66–75.
Georges, F.C. and Maynard, V.O. 1984. Separation of chromosomal DNA molecules from yeast by orthogonal-field-alternation gel electrophoresis.Nucleic Acid Res. 12: 5647–5664.
Gill, P., Jeffreys, A.J. and Werrett, D.J. 1985. Forensic application of DNA “fingerprints.”Nature 318: 577–579.
Jinks, D.C., Minter, M., Tarver, D.A., Vanderford, M., Hejtmancik, J.F. and McCabe, E.R.B. 1989. Molecular genetic diagnosis of sickle cell disease using dried blood specimens on blotters used for newborn screening.Hum. Genet. 81: 363–366.
Li, H., Gyllensten, U.B., Cui, X., Saiki, R.K., Erlich, H.A. and Arnheim, N. 1988. Amplification and analysis of DNA sequences in single human sperm and diploid cells.Nature 335: 414–417.
Maniatis, T., Fritsch, E.F. and Sambrook, J. 1982. InMolecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, pp. 149–186.
Matsumoto, T., Kondoh, T., Yoshimoto, M., Fujieda, K., Matsuura, N., Matsuda, I., Miike, T., Yano, K., Okuno, A., Aoki, Y., Murano, I., Toyota, S., Ohnishi, S. and Niikawa, N. 1988. Complex glycerol kinase deficiency: Molecular-genetic, cytogenetic, and clinical genetic studies of five Japanese patients.Am. J. Med. Genet. 31: 602–616.
McCabe, E.R.B., Huang, S.-Z., Selzer, W.K. and Law, M.L. 1987. DNA microextraction from dried blood spots on filter paper blotters: Potential applications to newborn screening.Hum. Genet. 75: 213–216.
Southern, E. 1975. Detection of specific sequences among DNA fragments separated by gel electrophoresis.J. Mol. Biol. 98: 503–517.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Hirota, T., Kondoh, T., Matsumoto, T. et al. Micro extraction of DNA from whole blood and amniocytes. Jap J Human Genet 34, 217–223 (1989). https://doi.org/10.1007/BF01900724
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/BF01900724