Particle Arrays 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 array comprising a population of particles disposed in a plurality of reaction chambers, wherein the particles each individually comprises one or more polynucleotides attached to a non-nucleosidic polymer network, and wherein at least one reaction chamber comprises, or is capacitively coupled to, a field effect transistor (FET). 
     
     
         25 . The array of  claim 24 , wherein at least one of the reaction chambers has a volume of no greater than about 10 μm 3 . 
     
     
         26 . The array of  claim 24 , wherein each particle in the population of particles has a volume, and the coefficient of variation of the volumes in the population of particles is no greater than about 15 percent. 
     
     
         27 . The array of  claim 24 , wherein at least one particle in the population of particles includes a clonal population of a single polynucleotide. 
     
     
         28 . The array of  claim 24 , wherein two or more particles in the population of particles each comprises a polynucleotide having a sequence that differs from the sequence of a polynucleotide of any other particle in the population. 
     
     
         29 . The array of  claim 24 , wherein at least one of the particles of the population includes a plurality of polynucleotides distributed throughout the volume of at least one particle, and wherein the polynucleotides are present at an average density of at least about 6.9×10 4  per μm 3 . 
     
     
         30 . The array of  claim 24 , wherein at least one of the particles in the population includes a plurality of polynucleotides having an average nearest neighbor distance of from about 15 to about 22 nm. 
     
     
         31 . The array of  claim 24 , wherein a plurality of particles in the population of particles have an average pore size of about 20 to about 150 nm. 
     
     
         32 . The array of  claim 24 , wherein a plurality the particles are substantially spherical or spheroidal in shape. 
     
     
         33 . The array of claim  9 , wherein a plurality of particles have diameters of from about 0.5 μm to about 10 μm. 
     
     
         34 . The array of  claim 24 , wherein a plurality of particles are non-spherical. 
     
     
         35 . The array of  claim 24 , wherein at least one of the field effect transistors of the array is an ion-sensitive field effect transistor (ISFET). 
     
     
         36 . The array of  claim 24 , wherein at least one of the plurality of reaction chambers includes no more than one particle.