US 6,270,970 B1 · Mixed-bed solid phase and its use in the isolation of nucleic acids

Abstract

Mixed-bed solid phases are provided, with methods for using such solid phases to isolate target nucleic acids, such as plasmid DNA, chromosomal DNA, RNA, or nucleic acids generated by enzymatic amplification from contaminants, including proteins, lipids, cellular debris, or other nucleic acids. The mixed-bed solid phases of this invention are mixtures of at least two different solid phases, each of which has a capacity to bind to the target nucleic acid under different solution conditions, and the capacity to release the nucleic acid under similar elution conditions. By exchanging solution conditions according to the methods of this invention, one can remove contaminants from the target nucleic acid bound to the mixed-bed solid phase, then elute the target nucleic acid in an elution buffer.

Claims19

Dependent claims

A mixed-bed solid phase for isolating a target nucleic acid from a mixture comprising the target nucleic acid and at least one contaminant, the mixed-bed solid phase comprising:

a first ion-exchanger solid phase having a capacity to bind to the target nucleic acid when combined with the mixture in the presence of a first solution having a first pH, and a capacity to release target nucleic acid bound thereto in the presence of a second solution having a second pH, wherein the first pH is at least 0.5 pH units different from the second pH;
and
a second ion-exchanger solid phase having a capacity to bind to the target nucleic acid when combined with the mixture in the presence of the second solution, and a capacity to release the target nucleic acid bound thereto in the presence of the first solution,
wherein the first ion-exchanger solid phase and second ion-exchanger solid phase are mixed together in a single container, thereby forming a mixed-bed solid phase, the first ion-exchanger solid phase and the second ion-exchanger solid phase of the mixed-bed solid phase each having a capacity to release the target nucleic acid bound thereto in the presence of an elution buffer.

1 more element

Dependencies are inferred from the claim text.

Mixed-bed solid phase and its use in the isolation of nucleic acids

Life of this patent

Mixed-bed solid phase and its use in the isolation of nucleic acids

Life of this patent

Abstract

Claims19