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Table 1 Intra- and inter-run variability of real-time PCR reactions. Intra- and inter-run variability was assayed for 5% RRS standard reference material, diluted to contain approximately 100, 20, 5, 1 and 0.5 ng of DNA. (Ct = Cycle threshold, SD = standard deviation of estimated copy number, CV = coefficient of variation of estimated copy number [%],* measurements outside the quantification range of the method)

From: Critical points of DNA quantification by real-time PCR – effects of DNA extraction method and sample matrix on quantification of genetically modified organisms

  

intra-assay

inter-assay

CV attributed to final result

  

Mean Ct

Copy number estimate

Mean Ct

Copy number estimate

 
 

Amount of DNA

 

Mean

SD

CV

 

Mean

SD

CV

 

lectin

100 ng (87000 copies)

22.12

76459

2652.86

3.47

22.21

76665

2136.80

2.79

7.3

 

20 ng (17400 copies)

24.26

19179

514.26

2.68

24.26

19561

562.87

2.88

4.4

 

5 ng (4350 copies)

26.38

4861

415.49

8.55

26.39

4705

145.13

3.08

12.5

 

1 ng (870 copies)

28.98

895

32.08

3.58

28.89

884

38.35

4.34

10.4

 

0.5 ng (435 copies)

30.25

392

20.92

5.33

30.06

403

13.71

3.40

10.9

RRS

100 ng (4350 copies)

27.99

4349

87.08

2.00

28.00

4017

273.05

6.80

 
 

20 ng (870 copies)

30.46

827

70.50

8.52

30.34

893

29.61

3.31

 
 

5 ng (218 copies)

32.39

224

23.36

10.41

32.51

223

27.00

12.10

 
 

1 ng (44 copies)

34.73

47

8.48

18.13

34.92

49

4.69

9.49

 
 

0.5 ng (22 copies)

35.90*

22

5.84

27.17

36.18*

22

2.30

10.34