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Methods and compositions for enriching compositions for polymerase enzyme complexes

  • US 10,544,457 B2
  • Filed: 06/09/2017
  • Issued: 01/28/2020
  • Est. Priority Date: 06/14/2016
  • Status: Active Grant
First Claim
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1. A method for enriching a loading composition for polymerase enzyme complexes, the method comprising:

  • (a) providing a loading composition comprising free polymerases and polymerase enzyme complexes, wherein the polymerase enzyme complexes are in an inactive form, and wherein each polymerase enzyme complex comprises a polymerase enzyme bound to a nucleic acid template;

    (b) applying a protease to the loading composition, which protease is more effective against the free polymerases than against polymerases in the polymerase enzyme complexes, and performing limited proteolytic digestion to reduce the amount of free polymerase relative to the amount of polymerase enzyme complex present in the composition, thereby enriching the loading composition for polymerase enzyme complexes; and

    (c) distributing the loading composition onto a substrate to provide a loaded substrate.

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