Materials and methods for the purification of polyelectrolytes, particularly nucleic acids
OwnerFolim G. Halakainventor
Life of this patent
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Abstract
This invention describes materials and methods usable for the purification of polyelectrolytes, such as nucleic acids and proteins. The materials of the invention are separation media that possess pH-dependent groups with pKa value in the range of about 5 to about 7. Separation of the nucleic acids or proteins from a separation medium is effected at a neutral or higher pH.
Claims9
A method for recovery of a DNA polynucleotide from a liquid containing said polynucleotide using a polymeric separation medium which contains 4-picolyl chloride pendant groups exhibiting a pKa value of less than about 7, which method comprises:
- (a) adjusting the pH of the liquid containing the polynucleotide to at least neutral pH and above the pKa of the pendant groups;
- (b) contacting the liquid containing the polynucleotide sample with the polymeric separation medium for a time period sufficient to cause the polynucleotide to form a water-insoluble polynucleotide-separation medium composite;
- (c) separating the liquid from the polynucleotide-separation medium composite;
- (d) washing the polynucleotide-separation medium composite with a buffer solution having at least neutral pH and above the pKa of the pendant groups, and;
- (e) contacting the separate composite with a buffer solution having at least neutral pH and at least 2 pH units above the pKa value of the pendant groups for a period of time sufficient to release the polynucleotide from separation medium.
A method for extracting DNA from microorganisms and biological cells present in a biological sample, which method comprises:
- (a) adding a nuclease inactivating substance to said sample in an amount sufficient to inhibit DNA degradation;
- (b) lysing the microorganisms and biological cells in said sample to release the DNA;
- (c) adjusting the pH of the lysed sample to a at least neutral and within about 2 pH units of the pKa of the pendant groups present in step (d);
- (d) contacting the lysed sample with a polymeric separation medium which contains 4-picolyl chloride pendant groups exhibiting a pKa value of less than about 7 for a time period sufficient to cause the DNA to form a DNA-separation medium composite;
- (e) separating the liquid from the DNA-separation medium composite, and;
- (f) contacting the DNA-separation medium composite with a buffer solution having at least neutral pH value and at least 2 units above the pKa value of the pendant groups for a period of time sufficient to release the nucleic acid from separation medium.
Dependencies are inferred from the claim text.