Table 1 Reaction inhibition, fluorescent signal, and melting temperature data: 16S amplicon
From: SYTO dyes and EvaGreen outperform SYBR Green in real-time PCR
Dye | Optimal Concentration | C t : 0.64 μM (0.32×) | C t : 2 μM (1×) | C t : 10 μM (5×) | C t : 20 μM (10×) | Maximum Fluorescence | T m shift slope (R 2 ) |
|---|---|---|---|---|---|---|---|
SYBR Green* | 0.64 μM (0.32×) | 27.4 ± 0.78 | no Ct | no Ct | no Ct | 1.00 | 5.89, × conc. 2.95, μM conc. (0.86) |
EvaGreen* | 5× | 31.54 ± 0.21 | 29.98 ± 0.50 | 28.60 ± 0.50 | not determined | 1.03 | 0.50 (0.94) |
Green* | Â | Â | Â | Â | Â | Â | Â |
SYTO 11 | 2 μM | 30.8 ± 1.26 | 29.9 ± 1.06 | 33.7 ± 2.58 | no Ct | 0.83 | 0.41 (0.96) |
SYTO 13 | 10 μM | 31.8 ± 1.06 | 30.5 ± 1.16 | 29.1 ± 0.95 | 29.9 ± 1.11 | 0.85 | 0.15 (0.92) |
SYTO 16 | 10 μM | 29.8 ± 0.95 | 28.5 ± 0.92 | 27.7 ± 1.04 | 30.3 ± 1.47 | 0.99 | 0.17 (0.93) |
SYTO 21 | 0.64 μM | 28.5 ± 0.98 | 32.94 ± 3.7 | no Ct | no Ct | 0.79 | 1.73 (0.63) |
SYTO 24 | 0.64 μM | 31.0 ± 0.0 | no Ct | no Ct | no Ct | 0.98 | not determined |
Orange* | Â | Â | Â | Â | Â | Â | Â |
SYTO 80 | 20 μM | 33.9 ± 0.57 | 32.4 ± 0.09 | 30.27 ± 0.01 | 29.4 ± 0.09 | 0.54 | 0.018 (0.33) |
SYTO 81 | 20 μM | 40.13 ± 0.0 | 35.7 ± 0.55 | 32.5 ± 0.10 | 32.0 ± 0.03 | 0.13 | -0.002 (0.003) |
SYTO 82 | 20 μM | 31.8 ± 0.37 | 31.25 ± 0.06 | 29.4 ± 0.01 | 28.7 ± 0.08 | 0.77 | 0.040 (0.59) |
SYTO 83 | 20 μM | 39.6 ± 2.09 | 35.2 ± 0.77 | 32.7 ± 0.41 | 32.4 ± 0.17 | 0.12 | 0.029 (0.38) |
Red** | Â | Â | Â | Â | Â | Â | Â |
SYTO 17 | 20 μM | no Ct | no Ct | 32.9 | 30.9 | 0.05 | -0.001 (0.009) |
SYTO 59 | 2 μM | 28.9 | 28.0 | 28.5 | 37.2 | 0.29 | 0.16 (0.96) |
SYTO 60 | 2 μM | 29.5 | 29.37 | no Ct | no Ct | 0.16 | 0.24 (0.78) |
SYTO 61 | 10 μM | 32.0 | 29.86 | 27.76 | 31.55 | 0.17 | 0.06 (0.91) |
SYTO 62 | 0.64 μM | 28.8 | 30.75 | no Ct | no Ct | 0.18 | 0.26 (0.99) |
SYTO 63 | 2 μM | 28.9 | 28.6 | 43.0 | no Ct | 0.20 | 0.12 (0.32) |
SYTO 64 | 2 μM | 34.8 | 31.7 | 31.7 | 34.0 | 0.15 | 0.012 (0.27) |