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Solid phase isothermal amplification

  • US 10,443,094 B2
  • Filed: 01/30/2017
  • Issued: 10/15/2019
  • Est. Priority Date: 03/26/2014
  • Status: Active Grant
First Claim
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1. A method for amplifying a nucleic acid, comprising:

  • a) providing a target nucleic acid template on a solid support;

    b) contacting the target nucleic acid template with a reaction mixture comprising a DNA polymerase having a strand displacement activity, a deoxyribonucleoside triphosphate (dNTP) mixture, a primer with a 3′

    end and a 5′

    end, a molecular crowding reagent, and a buffer solution, wherein the buffer solution maintains a salt concentration of the reaction mixture between 1 to 75 mM; and

    c) amplifying the target nucleic acid template under isothermal amplification conditions at a constant reaction temperature,wherein the salt concentration results in a melting temperature (Tm) of the primer at least 10°

    C. below the reaction temperature.

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