Scaffolded Nucleic Acid Polymer Particles and Methods of Making and Using
The invention provides particle compositions having applications in nucleic acid analysis. Nucleic acid polymer particles of the invention allow polynucleotides to be attached throughout their volumes for higher loading capacities than those achievable solely with surface attachment. In one aspect, nucleic acid polymer particles of the invention comprise polyacrylamide particles with uniform size distributions having low coefficients of variations, which result in reduced particle-to-particle variation in analytical assays. Such particle compositions are used in various amplification reactions to make amplicon libraries from nucleic acid fragment libraries.
1 .- 23 . (canceled)
24 . A particle comprising a non-nucleosidic polymer network and polynucleotides attached to the non-nucleosidic polymer network, wherein a plurality of the polynucleotides are attached to the network throughout its volume at a density of at least about 6.9×10 4 per μm 3 .
25 . The nucleic acid polymer particle of claim 1 , wherein the non-nucleosidic polymer network comprises agarose, polyoxybutylene, diethylacrylamide, polyoxyethylene, polyacrylamide, polyoxypropylene, N,N-polydimethylacrylamide, poly(N-isopropylacrylamide), polyvinylpyrrolidone, or poly-N-hydroxyacrylamide.
26 . The nucleic acid polymer particle of claim 2 , wherein the non-nucleosidic polymer network comprises polyacrylamide.
27 . The nucleic acid polymer particle of claim 3 , wherein the polyacrylamide has a T value of from about 5% to about 10% and a C value of from about 5% to about 10%.
28 . The nucleic acid polymer particle of claim 1 , wherein the plurality of polynucleotides are covalently attached to the non-nucleosidic polymer network throughout its volume at a density of at least about 1×10 5 per μm 3 .
29 . The nucleic acid polymer particle of claim 1 , wherein the plurality of polynucleotides are covalently attached to the non-nucleosidic polymer network via an acrydite group.
30 . The nucleic acid polymer particle of claim 1 , wherein the plurality of polynucleotides include a clonal population of polynucleotides.
31 . The nucleic acid polymer particle of claim 1 , wherein the nucleic acid polymer particle has an average pore size of from about 20 to about 150 nm.
32 . The nucleic acid polymer particle of claim 1 , wherein the nucleic acid polymer particle is substantially spherical or spheroidal in shape.
33 . The nucleic acid polymer particle of claim 32 , wherein the nucleic acid polymer particle has a diameter of from about 0.5 μm to about 30 μm.
34 . The nucleic acid polymer particle of claim 1 , wherein the nucleic acid polymer particle is non-spherical in shape.
35 . The nucleic acid polymer particle of claim 1 , wherein the nucleic acid polymer particle is stained with a dye.
36 . The nucleic acid polymer particle of claim 35 , wherein the dye is a fluorescent dye.
37 . The nucleic acid polymer particle of claim 1 , wherein a plurality of the polynucleotides have a length of between about 100 nucleotides and about 1200 nucleotides.