Polynucleotide Capture Materials, and Methods of Using Same
First Claim
Patent Images
1. A method for isolating polynucleotides from a cell-containing sample, the method comprising:
- contacting the sample with a lysis solution and a plurality of binding particles coated in PAMAM(Generation
0), so that the polynucleotides are liberated from the cells and become bound to the PAMAM(Generation
0), thereby creating binding particles bound with polynucleotides and a solution containing residual cellular matter;
compacting the binding particles bound with polynucleotides;
removing the solution containing residual cellular matter;
washing the binding particles; and
releasing the polynucleotides from the binding particles.
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Abstract
Methods for processing polynucleotide-containing biological samples, and materials for capturing polynucleotide molecules such as RNA and/or DNA from such samples. The RNA and/or DNA is captured by polyamidoamine (PAMAM(Generation 0)) bound to a surface, such as the surface of magnetic particles. The methods and materials have high efficiency of binding RNA and of DNA, and of release, and thereby permit quantitative determinations.
119 Citations
35 Claims
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1. A method for isolating polynucleotides 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 PAMAM(Generation
0), so that the polynucleotides are liberated from the cells and become bound to the PAMAM(Generation
0), thereby creating binding particles bound with polynucleotides and a solution containing residual cellular matter;compacting the binding particles bound with polynucleotides; removing the solution containing residual cellular matter; washing the binding particles; and releasing the polynucleotides from the binding particles. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 21, 22, 23, 26)
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27. A method for concentrating RNA from a sample containing polymerase chain reaction inhibitors, the method comprising:
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contacting between 500 μ
l and 1 ml of the sample with a plurality of RNA binding particles, the binding particles configured to preferentially retain the RNA in the sample as compared to the polymerase chain reaction inhibitors;concentrating the plurality of particles having the one or more polynucleotides bound thereto into an effective volume between 50 nanoliters and 5 microliters; and releasing the one or more polynucleotides into <
30 μ
l of solution.
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28. A composition comprising:
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carboxyl modified microparticles; and PAMAM(Generation
0) bound via one or more amine groups per molecule to one or more of the carboxylic acid groups on the microparticles.
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Specification