Methods and compositions for loading of polymerase complexes

Abstract

The present disclosure provides methods, compositions, and systems for distributing polymerase compositions into array regions. In particular, the described methods, compositions, and systems utilize density differentials and/or additives to increase efficiency in the distribution of polymerase compositions to a surface as compared to methods utilizing only diffusion control.

1 - 87 . (canceled) 
     
     
         88 . A method of distributing polymerase compositions onto a surface, the method comprising:
 (a) providing a polymerase composition in a solution comprising a stabilizing additive;   (b) adding the solution to the surface; and   (c) dehydrating the solution to concentrate the polymerase composition to the surface,   thereby distributing the polymerase composition to the surface.   
     
     
         89 . The method of  claim 111 , wherein the carbohydrate additive comprises a member selected from the group consisting of sucrose, dextrose, trehalose, and Ficoll. 
     
     
         90 . The method of  claim 88 , wherein the dehydrating step (c) comprises vacuum dehydration. 
     
     
         91 . The method of  claim 88 , wherein the surface comprises a plurality of array regions. 
     
     
         92 . The method of  claim 91 , wherein the plurality of array regions comprise nanowells. 
     
     
         93 . The method of  claim 88 , wherein the polymerase composition comprises polymerase-template complexes. 
     
     
         94 . The method of  claim 93 , wherein subsequent to the distributing, the polymerase-template complexes are processed to identify nucleotide sequences of the template. 
     
     
         95 . The method of  claim 88 , wherein subsequent to the dehydrating step (c), the polymerase composition is rehydrated with deionized water. 
     
     
         96 - 110 . (canceled) 
     
     
         111 . The method of  claim 88 , wherein the stabilizing additive is a carbohydrate additive. 
     
     
         112 . The method of  claim 111 , wherein the carbohydrate additive is present in the solution at a concentration of 5-70% weight/volume. 
     
     
         113 . The method of  claim 88 , wherein the stabilizing additive is polyethylene glycol (PEG). 
     
     
         114 . The method of  claim 113 , wherein the stabilizing additive is PEG-8000. 
     
     
         115 . The method of  claim 88 , wherein the stabilizing additive is sorbitol.