Figure 5 | Scientific Reports

Figure 5

From: Reduced False Positives and Improved Reporting of Loop-Mediated Isothermal Amplification using Quenched Fluorescent Primers

Figure 5

Detection with SYTO9 and JOE-FIP positive and negative samples. (1) LAMP amplification of 500000 copies linear plasmid template and NTCs with detection by both (1A) and JOE-FIP quenched fluorescence (1B) in separate tubes. Four replicates of template in blue and NTCs in red. Melt curve analysis (1C,D) showing amplification of template only in the 90 minute assay. (2) Amplification with LAMP primers of 72 replicate NTCs for 200 minutes. (2A) SYTO9 detection of amplification between 150 and 200 minutes showing 5 of 72 giving a positive signal. (2B) JOE-FIP detection of the same samples using the yellow channel (excitation at 530 +/− 5 nm with emission at 555 nm +/− 5 nm). (2C) FAM-LoopF detection using the green channel (excitation at 470 nm +/− 10 nm with emission at 510 nm +/− 5 nm). (2D–F) Melt curve analyses.

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