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Methods and compositions for single molecule composition loading

  • US 10,300,452 B2
  • Filed: 03/23/2016
  • Issued: 05/28/2019
  • Est. Priority Date: 03/24/2015
  • Status: Active Grant
First Claim
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1. A method of distributing single polymerase molecules into array regions, the method comprising;

  • (a) providing a surface comprising a plurality of array regions, wherein each array region comprises several binding elements;

    (b) exposing the surface to a solution comprising polymerase enzyme compositions, wherein each polymerase enzyme composition comprises a polymerase bound to a scaffold, wherein the scaffold comprises;

    (i) a core comprising a material other than DNA, and(ii) at least three linear arms comprising biotin moieties attached to DNA molecules, wherein each arm comprises multiple biotin moieties,wherein the exposing is conducted under conditions such that the biotin moieties of the scaffold react with available binding elements in a given array region and prevent other polymerase enzyme compositions from loading in that given array region,thereby distributing single polymerase molecules into array regions.

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