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. 2021 Sep;32(3):89–97. doi: 10.7171/jbt.21-3203-003

FIGURE 1.

FIGURE 1

Real-time amplification data demonstrate rapid and sensitive detection. A) Bst 2.0 is more rapid than Bst LF but also more prone to non-template amplification. N250/N1000 reactions have 250/1000 copies of the N plasmid control, respectively. Incubation temperature was at 64°C, and reaction volumes were 20 μl. B) RNA controls are reverse transcribed and amplified for detection down to a single copy per microliter of the reaction. Synthetic RNA (BEI Resources) dilutions were spiked into duplicate reactions at 20, 50, 100, and 200 (R20, R50, R100, and R200) copies, in addition to nontemplate control (NTC) and DNA controls. These reactions used Bst 2.0 and RTx; incubation included an initial 10 min at 55°C for reverse transcription before shifting up to 64°C for the real-time acquisition.RFU - relative fluorescence units.