Organic compounds -- part of the class 532-570 series – Organic compounds – Carbohydrates or derivatives
Patent
1996-11-13
1998-05-12
Saunders, David
Organic compounds -- part of the class 532-570 series
Organic compounds
Carbohydrates or derivatives
536127, C07H 2104, C07H 108
Patent
active
057506710
ABSTRACT:
This invention relates to a method for isolating DNA from biological cells containing protamines. The biological parent material is decomposed using salts with low molecular weight above a critical ionic strength cleaving all ionic bonds between DNA and protamines and polystyrene sulfonic acid (PSA) and/or its derivatives in combination with sodium lauryl sulfate. These substances, together with protamines at pH values ranging from 7.5 to 8.5, form insoluble polyelectrolyte complexes facilitating the release of DNA from its natural complexes which can then be precipitated using ethanol. The method according to the invention is fundamentally different from conventional methods as it uses neither enzymes nor toxic extracting agents such as chloroform or phenol. The method according to the invention is very efficient and non-polluting; it can be carried out easily and yields products with a high degree of purity. As the treatment is very gentle, this method is suitable to purify DNA of very high molecular weight.
REFERENCES:
Rodman, TC et al. Exp. Cell Res. 150(2):269-281, Feb. 1984.
Doenecke, Interaction of Polystyrene Sulfonate with Nucleosomes, Biochemistry International, vol. 1, No. 3, pp. 237-244, Sep. 21, 1980.
Tubbert et al., Chromatin and Histones in Mealy Bug Spermatogonia, Experimental Cell Research 85, pp. 205-211, 1974.
Trihn et al., Ionic Strength Dependence of the Stability of Polyelectrolyte Complexes, Die Angewandte Makromolekulare Chemie, vol. 212, No. 3718, pp. 167-179, (1993).
Sambrook et al., Molecular Cloning, (second edition) pp. 14-22, (1989).
Zahn et al., Ein Konservierungs-und Darstellungs-Verfahren fur Desoxyribonucleinsauren unde ihre Ausgangsmaterialien, Biochemische Zeitschrift, vol. 336, pp. 281-298, (1962).
Marmur, A Procedure for the Isolation of Deoxyribonucleic Acid from Micro-organisms, J. Mol. Biol., vol. 3, pp. 208-218, (1961).
Saunders David
VanderVegt F. Pierre
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