Selected article for: "copy number and specific amplification"
Author: Gun-Soo Park; Keunbon Ku; Seung-Hwa Beak; Seong Jun Kim; Seung Il Kim; Bum-Tae Kim; Jin-Soo Maeng
Title: Development of Reverse Transcription Loop-mediated Isothermal Amplification (RT-LAMP) Assays Targeting SARS-CoV-2
  • Document date: 2020_3_12
    • ID: 1qn5y4gc_23
    Snippet: To optimize RT-LAMP reaction with Bst3.0, we first evaluated optimal concentration of dNTP and Mg 2+ for each primer set. Three dNTP-Mg 2+ concentration combinations are tested at 69℃: dNTP 1.4 mM and Mg 2+ 8 mM, dNTP 1.4 mM and Mg 2+ 6 mM, dNTP 1 mM and Mg 2+ 6 mM. The copy number of RNA template was 1,000 copies per reaction. Best concentration combination was selected for each primer set by whether both of duplicate show specific amplificati.....
    Document: To optimize RT-LAMP reaction with Bst3.0, we first evaluated optimal concentration of dNTP and Mg 2+ for each primer set. Three dNTP-Mg 2+ concentration combinations are tested at 69℃: dNTP 1.4 mM and Mg 2+ 8 mM, dNTP 1.4 mM and Mg 2+ 6 mM, dNTP 1 mM and Mg 2+ 6 mM. The copy number of RNA template was 1,000 copies per reaction. Best concentration combination was selected for each primer set by whether both of duplicate show specific amplification and by threshold time. Average threshold time (Tt_av) and corresponding dNTP and Mg 2+ concentrations are as follows: dNTP 1 mM and Mg 2+ 6 mM for "Nsp3_1-61" (Tt_av = 11.93), dNTP 1.4 mM and Mg 2+ 8 mM for "Nsp3_2-24" (Tt_av = 7.74), dNTP 1.4mM and Mg 2+ 6mM for "S_1-2-2" and "N_21" (Tt_av = 11.08 and 5.50, respectively).

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