Particle Population 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 . An amplicon library comprising a plurality of particles, each particle including an amplicon and a porous non-nucleosidic polymer network, each amplicon including a plurality of polynucleotides that are attached to a non-nucleosidic polymer network throughout its volume at a density of at least about 6.9×10 4  per μm 3 , where at least one particle includes a polynucleotide having a sequence that is different from a polynucleotide of any other particle in the population. 
     
     
         25 . The amplicon library of  claim 24 , where the volume of at least one particle of the plurality of particles is less than about 1.4×10 4  μm 3 . 
     
     
         26 . The amplicon library of  claim 24 , wherein the volume of at least one particle of the plurality of particles is less than about 1.8×10 3  μm 3 . 
     
     
         27 . The amplicon library of  claim 24 , wherein at least one particle of the plurality of particles is spheroidal in shape. 
     
     
         28 . The amplicon library of  claim 24 , wherein the non-nucleosidic polymer network of at least one particle of the plurality of particles includes a polyacrylamide gel having a T value from 5% to 10% and a C value from 5% to 10%. 
     
     
         29 . The amplicon library of  claim 24 , wherein at least one particle of the plurality of particles includes a one or more polynucleotides having an average length of about 100 to about 1,200 nucleotides. 
     
     
         30 . The amplicon library of  claim 24 , wherein at least one particle of the plurality of particles includes one or more polynucleotides that are double stranded. 
     
     
         31 . The amplicon library of  claim 24 , wherein at least one particle of the plurality of particles includes one or more polynucleotides that are attached to the non-nucleosidic polymer network of the particle throughout the volume of the network at an average nearest neighbor distance of from about 15 to about 22 nm. 
     
     
         32 . The amplicon library of  claim 24 , wherein the coefficient of variation of the volumes of the plurality of particles is no greater than about 15 percent. 
     
     
         33 . The amplicon library of  claim 24 , wherein the plurality of particles are disposed on an array comprising a plurality of reaction chambers, wherein at least one reaction chamber comprises or is capacitively coupled to a field effect transistor (FET). 
     
     
         34 . The array of  claim 33 , wherein the field effect transistor is an ion-sensitive field effect transistor (ISFET).