POLYNUCLEOTIDE CAPTURE MATERIALS, AND SYSTEMS USING SAME
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 polyamindoamine (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.
1 . (canceled)
2 . A system comprising:
a container comprising:
a plurality of magnetic beads coated with polycationic polyamine dendrimers, the polycationic polyamine dendrimers being covalently bonded to the plurality of magnetic beads via amide bonds and comprising at least three dendrimer branches having primary amine terminals, and
a lytic enzyme,
wherein the container is configured to receive a biological sample containing polynucleotides and to allow the biological sample to come in contact with the plurality of magnetic beads, and
wherein the plurality of magnetic beads in the container are configured to reversibly bind polynucleotides of the biological sample, thereby producing a set of polynucleotide-retained beads;
a release solution, wherein the set of polynucleotide-retained beads are configured to release the polynucleotides when contacted with a release solution and heated.
3 . The system of claim 2 , wherein the plurality of magnetic beads and the lytic enzyme are in dry form in the container prior to receiving the biological sample in the container.
4 . The system of claim 2 , wherein the polycationic polyamine dendrimers comprise six amine groups prior to covalently bonding to the plurality of magnetic beads.
5 . The system of claim 2 , wherein the polycationic polyamine dendrimers have a molecular weight of less than about 600 Da prior to covalently bonding to the plurality of magnetic beads.
6 . The system of claim 2 , wherein the polycationic polyamine dendrimers comprise at least four dendrimer branches having primary amine terminals prior to covalently bonding to the plurality of magnetic beads.
7 . The system of claim 2 , wherein the polycationic polyamine dendrimers comprise at least five amine groups when covalently bonded to the plurality of magnetic beads.
8 . The system of claim 2 , wherein the at least one container further comprises sample process control polynucleotides in dry form prior to receiving the biological sample in the at least one container.
9 . The system of claim 2 , wherein the lytic enzyme comprises proteinase K.
10 . The system of claim 2 , wherein the polycationic polyamine dendrimers comprise an initial monomer core of ethylene diamine.
11 . A kit comprising:
a plurality of containers, at least one container comprising:
a plurality of magnetic beads coated with polycationic polyamine dendrimers, the polycationic polyamine dendrimers being covalently bonded to carboxyl groups on the plurality of magnetic beads, and
a lytic enzyme,
wherein the at least one container is configured to receive a biological sample containing polynucleotides and allow the biological sample to come in contact with the plurality of magnetic beads, and
wherein the plurality of magnetic beads in the at least one container are configured to reversibly bind polynucleotides of the biological sample, through ionic bonding with the polycationic polyamine dendrimers, thereby producing a set of polynucleotide-retained beads;
wherein the set of polynucleotide-retained beads are configured to release the polynucleotides when contacted with a release solution and heated.
12 . The kit of claim 11 , wherein the polycationic polyamine dendrimers comprise six amine groups prior to covalently bonding to carboxyl groups on the magnetic beads.
13 . The kit of claim 11 , wherein the polycationic polyamine dendrimers comprise at least four dendrimer branches having primary amine terminals prior to covalently bonding to carboxyl groups on the magnetic beads.
14 . The kit of claim 11 , wherein the polycationic polyamine dendrimers comprise at least five amine groups when covalently bonded to carboxyl groups on the magnetic beads.
15 . The kit of claim 11 , wherein the polycationic polyamine dendrimers comprise at least three dendrimer branches having primary amine terminals when covalently bonded to carboxyl groups on the magnetic beads.
16 . The kit of claim 11 , wherein the polycationic polyamine dendrimers are amide-bonded to the magnetic beads.
17 . The kit of claim 11 , wherein the polycationic polyamine dendrimers comprise polyamide dendrimers.
18 . The kit of claim 17 , wherein the polyamide dendrimers comprise PAMAM dendrimers.
19 . The kit of claim 11 , wherein the at least one container further comprises sample process control polynucleotides prior to receiving the biological sample in the container.
20 . The kit of claim 11 , wherein the lytic enzyme comprises proteinase K.
21 . The kit of claim 11 , wherein the polycationic polyamine dendrimers comprise an initial monomer core of ethylene diamine.