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Table 2 Reaction inhibition, fluorescent signal, and melting temperature data: tropomyosin 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

33.00 ± 0.10

35.07 ± 1.10

no Ct

no Ct

1.00

3.99, × conc.

1.99, μM conc.

(0.97)

EvaGreen*

5×

36.37 ± 0.48

35.04 ± 0.35

35.15 ± 1.13

not determined

0.91

0.36 (0.98)

Green*

       

SYTO 11

2 μM

38.02 ± 0.97

36.42 ± 0.84

38.17 ± 1.03

42.27 ± 0

0.51

-0.20 (0.12)

SYTO 13

10 μM

37.58 ± 1.33

36.10 ± 1.32

36.06 ± 1.55

36.45 ± 2.06

0.87

0.001 (0.00)

SYTO 16

10 μM

37.00 ± 1.86

34.20 ± 1.05

34.04 ± 0.96

34.97 ± 0.53

1.07

0.030 (0.004)

SYTO 21

0.64 μM

34.55 ± 1.07

35.90 ± 0.83

no Ct

no Ct

0.82

0.62 (0.03)

SYTO 24

0.64 μM

33.50 ± 0.01

no Ct

no Ct

no Ct

1.02

not determined

Orange*

       

SYTO 80

20 μM

39.16 ± 2.44

37.33 ± 1.86

35.84 ± 0.02

34.78 ± 0.08

0.40

0.048 (0.008)

SYTO 81

20 μM

43.89 ± 0.10

40.62 ± 1.39

37.02 ± 0.39

36.45 ± 0.21

0.11

0.026 (0.002)

SYTO 82

20 μM

37.18 ± 1.85

35.59 ± 0.95

33.50 ± 0.11

33.44 ± 0.51

0.61

-0.17 (0.10)

SYTO 83

20 μM

42.20 ± 0.01

39.28 ± 0.99

37.04 ± 0.13

35.92 ± 0.93

0.10

0.037 (0.0051)

Red*

       

SYTO 17

20 μM

no Ct

42.26 ± 0.02

36.18 ± 0.14

35.32 ± 0.41

0.03

0.0009 (0.0005)

SYTO 59

10 μM

35.63 ± 0.09

35.15 ± 1.50

33.70 ± 0.10

35.53 ± 0.53

0.20

0.11 (0.83)

SYTO 60

2 μM

34.85 ± 0.41

34.29 ± 0.31

41.09 ± 1.97

no Ct

0.13

0.064 (0.10)

SYTO 61

10 μM

37.45 ± 1.68

35.38 ± 0.57

34.14 ± 0.98

35.00 ± 1.19

0.14

0.043 (0.58)

SYTO 62

0.64 μM

33.98 ± 0.42

33.69 ± 0.75

no Ct

no Ct

0.17

0.035 (0.008)

SYTO 63

2 μM

34.78 ± 0.41

34.50 ± 0.31

37.07 ± 0.07

no Ct

0.12

0.24 (0.85)

SYTO 64

2 μM

36.22 ± 0.10

36.68 ± 1.62

36.31 ± 1.05

36.48 ± 0.95

0.05

-0.004 (0.013)

  1. No fluorescent signals were obtained for SYTO dyes 12, 14, 25, 84, or 85
  2. *Ct values are average ± SD for at 2-3 independent experiments