Figure 2
Specificity, melting peak and standard curve analysis of the SYBR Green qPCR assay with the Bam249F/R and BS310F/R primer sets. (A) B. velezensis. (a) Fluorescence intensity as a function of template concentration. For each assay, a series of 10-fold dilutions of cloned DNA (ranging from 1.42 × 103 to 1.42 × 109 copies/µl) was used as the template (1–7, sample dilutions). (b) Standard curve derived from the amplification plot. (c) Melting curve analysis (1–7, sample dilutions). (d) Melting peak analysis (1–7, sample dilutions). The amplified product derivatives of the relative fluorescence units [-d(RFU)/dT] were plotted as a function of temperature (amplified product, 86.0 °C). The large peak indicates the amplified product, while the small peak indicates the no-template control. (B) B. subtilis subsp. subtilis. (a) Fluorescence intensity as a function of template concentration. For each assay, a series of 10-fold dilutions of cloned DNA (ranging from 1.39 × 103 to 1.39 × 109 copies/µl) was used as the template (1–7, sample dilutions). (b) Standard curve derived from the amplification plot. (c) Melting curve analysis (1–7, sample dilutions). (d) Melting peak analysis (1–7, sample dilutions). The amplified product derivatives of the relative fluorescence units [-d(RFU)/dT] were plotted as a function of temperature (amplified product, 83.5 °C). The large peak indicates the amplified product, while the small peak indicates the no-template control.
