SOLID PHASE ISOTHERMAL AMPLIFICATION

• US 20170137874A1
• Filed: 01/30/2017
• Published: 05/18/2017
• Est. Priority Date: 03/26/2014
• Status: Active Grant

• Current Assignee: Global Life Sciences Solutions Operations UK Ltd. (Danaher Corporation)
• Original Assignee: General Electric Company
• Inventors: Horton, Jeffrey Kenneth, Heller, Ryan Charles, Nelson, John Richard, Patel, Paresh Lakhubhai, Wakefield, Alison Myfanwy, Capper, Stephen James, Tatnell, Peter James
• Granted Patent: US 10,443,094 B2
• Time in Patent Office: Days
• CPC Class Codes: C12P 19/34 Polynucleotides, e.g. nucleic acids, oligoribonucleotides
• CPC Class Codes: C12Q 1/6844 Nucleic acid amplification reactions
• CPC Class Codes: C12Q 2527/107 Temperature of melting, i.e. Tm
• CPC Class Codes: C12Q 2527/125 Specific component of sample, medium or buffer
• CPC Class Codes: C12Q 2527/137 Concentration of a component of medium
• CPC Class Codes: C12Q 2531/119 Strand displacement amplification [SDA]

Abstract

Provided herein are methods and kits for isothermal nucleic acid amplifications that use a target nucleic acid template; 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 for amplifying the target nucleic acid template. The buffer solution maintains a low salt concentration of the reaction mixture, and wherein the salt concentration results in a melting temperature (T_m) of the primer at least 10° C. below the reaction temperature. The amplification is effected under isothermal condition.

Claims