Polynucleotide Capture Materials, and Method of Using Same
First Claim
Patent Images
1. A method for isolating DNA from a cell-containing sample, the method comprising:
- contacting the sample with a lysis solution and a plurality of binding particles coated in polyethyleneimine, so that the DNA is liberated from the cells and becomes reversibly bound to the polyethyleneimine, thereby creating binding particles bound with DNA and a solution containing residual cellular matter;
compacting the binding particles bound with DNA;
removing the solution containing residual cellular matter;
washing the binding particles; and
releasing the DNA from the binding particles.
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Abstract
Methods for processing polynucleotide-containing biological samples, and materials for capturing polynucleotide molecules such as DNA from such samples. The DNA is captured by polyethyeleneimine (PEI) bound to a surface, such as the surface of magnetic particles. The methods and materials have high efficiency of binding DNA and of release, and thereby permit quantitative determinations.
227 Citations
57 Claims
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1. A method for isolating DNA from a cell-containing sample, the method comprising:
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contacting the sample with a lysis solution and a plurality of binding particles coated in polyethyleneimine, so that the DNA is liberated from the cells and becomes reversibly bound to the polyethyleneimine, thereby creating binding particles bound with DNA and a solution containing residual cellular matter; compacting the binding particles bound with DNA; removing the solution containing residual cellular matter; washing the binding particles; and releasing the DNA from the binding particles. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 13, 14, 18, 19, 20, 29, 30, 32, 33, 34, 38, 39, 43)
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- 40. The method of claim 40, wherein after removal of the solution containing residual cellular matter, less than 10 microliters of solution is left along with the particles.
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51. A kit, comprising:
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a number of sealed tubes, each containing lysis buffer; a tube containing lyophilized microparticles having polyethyeleneimine bound thereto; a tube containing liquid wash reagents, sufficient to analyze the number of samples; a tube containing liquid neutralization reagents, sufficient to analyze the number of samples; and a tube containing liquid release reagents, sufficient to analyze the number of samples, wherein each component of the kit is stored in an air-tight container.
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54. A kit, comprising:
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a first air-tight pouch enclosing a number of tubes, each tube containing lyophilized microparticles having polyethyeleneimine bound thereto; a second air-tight pouch enclosing a number of reagent holders, each holder comprising; a tube containing liquid lysis reagents; a tube containing liquid wash reagents; a tube containing liquid neutralization reagents; and a tube containing liquid release reagents.
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Specification