Method for using aqueous two-phase system for the isolation, purification and/or concentration of short nucleic acid fragments
AssigneePhase Scientific International, Ltd.
InventorYin To CHIU
Life of this patent
Estimating the term…
Abstract
The present invention relates to methods of using aqueous two-phase system (ATPS) for the isolation, concentration and/or purification of short nucleic acid fragment having about or less than 250 base pairs (bp). In one embodiment, the present invention provides a composition and kit for the purification of short nucleic acid fragments having about or less than 250 base pairs from nucleic acid-containing biological materials. In another embodiment, the present invention provides uses of certain salts and/or polymers in a two-phase system for the purification of short nucleic acid fragments having about or less than 250 base pairs from nucleic acid containing biological materials.
Claims17
A method for selectively isolating and concentrating nucleic acid fragments of 250 bp or less from a sample, comprising:
- a) preparing a first composition comprising a first phase component and a second phase component forming a first aqueous two phase system (ATPS) when the components are dissolved in an aqueous solution, wherein the first phase component is a polymer dissolving in a first phase, and the second phase component is a salt dissolving in a second phase;
- b) mixing the composition from step a) with an aqueous solution containing said sample, allowing the resulting mixture to partition into two phases, wherein said nucleic acid fragments are concentrated in the bottom phase;
- c) isolating the bottom phase;
- d) preparing a second composition comprising a third phase component and a fourth phase component forming a second aqueous two phase system (ATPS) when the components are dissolved in an aqueous solution, wherein the third phase component is a polymer dissolving in a third phase, and the fourth phase component is a salt dissolving in a fourth phase;
- e) mixing the composition from step d) with the bottom phase of step c), allowing the resulting mixture to partition into two phases, wherein said nucleic acid fragments are concentrated in the top phase;
- f) isolating the top phase that is concentrated with said nucleic acid fragments of 250 bp or less.
A kit for selectively isolating and concentrating nucleic acid fragments of 250 bp or less from a sample, characterized in that the kit comprises:
- (a) a first composition comprising a first phase component and a second phase component forming a first aqueous two phase system (ATPS) when the components are dissolved in an aqueous solution, wherein the first phase component is 10-30% (w/w) polyethylene glycol 1000 and the second phase component is 5-20% (w/w) potassium phosphate dibasic;
- and
- (b) a second composition comprising a third phase component and a fourth phase component forming a second aqueous two phase system (ATPS) when the components are dissolved in an aqueous solution, wherein the third phase component is 5-20% (w/w) polyethylene glycol 6000 and the fourth phase component is 15-25% (w/w) potassium phosphate dibasic.
Dependencies are inferred from the claim text.