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Table 3 Comparison of simultaneous analysis and standard curve approach. Data for table 2 was analyzed as 3 subgroups of 5 samples each. In the standard curve approach, the EA was calculated as 10-1/slope with the slope of the regression line. Relative error of accuracy was calculated as the absolute value of 1 – (measured factor/4).

From: Simultaneous fitting of real-time PCR data with efficiency of amplification modeled as Gaussian function of target fluorescence

  

Standard curve (Pfaffl) approach

Present paper approach

Target

Dilution step 1 : 4

EA subgroup

Factor arithmetic mean

SEM

Precision relative error (%)

Accuracy relative error (%)

EA0 subgroup

Factor arithmetic mean

SEM

Precision relative error (%)

Accuracy relative error (%)

SLC6A14 rat

1

1.77

4.8

0.5

11

21

1.92

4.5

0.2

3

14

 

2

1.74

5.0

1.4

28

25

1.92

5.7

1.0

17

42

 

3

1.80

3.1

0.9

28

21

1.89

3.6

0.5

14

9

 

4

 

4.7

1.3

28

18

 

4.9

1.1

23

22

SLC22A13 human

1

2.08

2.9

0.2

7

29

1.86

3.6

0.1

3

10

 

2

2.04

5.7

0.2

3

43

1.86

4.3

0.1

3

6

 

3

1.82

2.7

0.3

12

33

1.86

3.3

0.2

6

17

 

4

 

6.5

1.4

21

61

 

3.7

0.4

11

8

EMT pig

1

1.71

3.1

0.1

4

23

1.81

3.3

0.1

2

18

 

2

1.76

2.5

0.9

35

38

1.79

4.1

0.3

8

3

 

3

1.77

9.6

4.1

42

140

1.78

7.0

3.5

50

75

 

4

 

5.3

1.8

33

33

 

4.2

1.2

28

5

ETT chicken

1

1.87

6.7

2.0

30

66

1.89

5.3

1.5

27

33

 

2

2.01

2.8

0.6

22

31

1.89

2.9

0.7

25

28

 

3

1.83

4.9

0.6

12

21

1.86

4.3

0.3

8

7

 

4

 

3.7

0.4

10

7

 

3.9

0.2

5

3

GAPDH human

1

1.91

2.0

0.1

5

50

2.00

3.0

0.1

3

26

 

2

1.92

5.0

0.3

6

26

1.96

4.0

0.3

7

0

 

3

1.84

4.9

0.1

1

22

1.99

4.8

0.2

5

20

 

4

 

4.3

0.1

2

6

 

4.5

0.4

9

12

    

Median:

12

27.5

  

Median:

8

13