Polynucleotide capture materials, and methods of using same

摘要

Disclosed are methods for isolating polynucleotides (e.g., RNA and DNA) from a cell-containing sample. The disclosed methods employ polyamidoamine (PAMAM) dendrimer molecules bound to the surface of binding particles to capture and isolate the polynucleotides from the samples. The polynucleotides are released from the binding particles by contacting the binding particles with a release solution having a basic pH.

主权项

1. A method for isolating polynucleotides from a cell-containing sample in a process tube, the method comprising:
lysing the cells in the sample by contacting the sample with a lysis solution having a pH between 4 and 8, thereby releasing polynucleotides from the cells; retaining the polynucleotides in the sample as compared to any amplification inhibitors by contacting the sample with magnetic binding particles having PAMAM(Generation 0) molecules covalently bound to the surface thereof, the polynucleotides becoming reversibly retained by the PAMAM(Generation 0) molecules on the binding particles; compacting the binding particles retaining the polynucleotides in the process tube by positioning a magnet in communication with the binding particles; removing the lysis solution from the process tube; washing the binding particles with a wash solution having a pH less than 9 while the polynucleotides are retained on the binding particles; and releasing the polynucleotides from the binding particles by contacting the binding particles with a release solution having a pH between 9 and 12.