Immobilized Buffer Particles and Uses Thereof

Abstract

The disclosure relates to novel particle compositions and methods of making said compositions having applications in nucleic acid analysis, as well as apparatuses and systems for the same.

1 .- 74 . (canceled) 
     
     
         75 . An apparatus comprising:
 a volume defined by at least one surface having one or more confinement regions, the volume containing one or more immobilized buffer particles including a polymer network linked to one or more buffering groups, the one or more immobilized buffer particles having an average size and shape such that substantially no immobilized buffer particles can enter the one or more confinement regions; and   a pH sensor operationally associated with at least one of the one or more confinement regions, wherein the pH sensor is configured to measure a pH of the reagent in the at least one confinement region.   
     
     
         76 . The apparatus of claim  68 , wherein the polymer network comprises at least one polymer that is hydrophilic. 
     
     
         77 . The apparatus of claim  68 , wherein the polymer network includes a network structure having one or more pores, wherein at least one of the one or more pores permits access to the interior of particle by solvents or reagents. 
     
     
         78 . The apparatus of claim  68 , wherein at least one of the one or more buffering groups is selected from the group consisting of triethanolamine, N-[tris(hydroxymethyl)methyl]-2-aminoethanesulfonic acid, 3-(N-tris[hydroxymethyl]methylamino)-2-hydroxypropanesulfonic acid, N-(2-hydroxyethyl)piperazine-N-(2-ethanesulfonic acid), N-(2-acetamido)-2-aminoethanesulfonic acid, imidazole, and acetate. 
     
     
         79 . The apparatus of claim  68 , wherein at least one of the one or more buffering groups has a pKa within the range of about 4.5 to about 9.0. 
     
     
         80 . The apparatus of  claim 79 , wherein the at least one buffering group has a pKa in the range of about 6.0 to about 8.0. 
     
     
         81 . The apparatus of claim  68 , wherein at least one of the one or more immobilized buffer particles includes an electrically charged particle. 
     
     
         82 . The apparatus of  claim 81 , wherein the electrically charged particle is situated inside an apparatus for measuring pH values that includes a confinement region containing one or more zones of like charge, and wherein the electrically charged particle has an electrical charge having the same polarity as the like charges contained in the one or more zones. 
     
     
         83 . The apparatus of claim  68 , wherein at least one of the one or more immobilized buffer particles has a minimum length in any one dimension that is greater than the diameter of at least one of the one or more confinement region. 
     
     
         84 . The apparatus of claim  68 , wherein the polymer network of at least one of the one or more immobilized buffer particles includes a network structure having one or more pores, wherein at least one of the one or more pores permits access to the interior of the at least one immobilized buffer particle by solvents or reagents. 
     
     
         85 . The apparatus of claim  68 , wherein at least one of the plurality of immobilized buffer particles has a size and wherein the plurality of immobilized buffer particles has a size distribution with a coefficient of variation of less than about 50%. 
     
     
         86 . The apparatus of claim  68 , wherein the pH sensor includes an ISFET. 
     
     
         87 . The apparatus of claim  68 , wherein the average diameter of the one or more immobilized buffer particles is greater than the average diameter of the one or more confinement regions. 
     
     
         88 . The apparatus of claim  68 , wherein at least one of the one or more immobilized buffer particles includes a spheroidal polyacrylamide particle.