Scaffolded Nucleic Acid Polymer Particles and Methods of Making and Using

Abstract

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-nucleosidie polymer network and polynucleotides attached to the non-nucleosidie polymer network throughout its volume, wherein a plurality of the polynucleotides are attached to the network at an average nearest neighbor distance of from about 15 to about 22 nm. 
     
     
         25 . The particle of  claim 24 , wherein the plurality of polynucleotides are covalently attached to the non-nucleosidie polymer network throughout its volume at an average nearest neighbor distance of from about 18 to about 20 nm. 
     
     
         26 . The particle of  claim 24 , wherein the non-nucleosidie polymer network comprises agarose, polyoxybutylene, diethylacrylamide, polyoxyethylene, polyacrylamide, polyoxypropylene, N,N-polydimethylacrylamide, poly(N-isopropylacrylamide), polyvinylpyrrolidone, or poly-N-hydroxyacrylamide. 
     
     
         27 . The particle of  claim 26 , wherein the non-nucleosidie polymer network comprises polyacrylamide. 
     
     
         28 . The particle of  claim 27 , 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%. 
     
     
         29 . The particle of  claim 25 , wherein the polynucleotides are covalently attached to the non-nucleosidic polymer network via an acrydite group. 
     
     
         30 . The particle of  claim 24 , wherein the plurality of the polynucleotides include a clonal population of polynucleotides. 
     
     
         31 . The particle of  claim 24 , wherein the particle has an average pore size of from about 20 to about 150 nm. 
     
     
         32 . The particle of  claim 24 , wherein the particle is a spherical particle having a diameter of from about 0.5 μm to about 10 μm. 
     
     
         33 . The particle of  claim 24 , wherein the particle is stained with a dye. 
     
     
         34 . The particle of  claim 33 , wherein the dye is a fluorescent dye. 
     
     
         35 . The particle of  claim 24 , wherein the polynucleotides have a length of 100-1,200 nucleotides.