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Table 2 RNase If -qPCR assay inter-assay precision

From: RNase If -treated quantitative PCR for dsRNA quantitation of RNAi trait in genetically modified crops

 

dsRNA input for qPCR (pg)

Run 1 Cp

Run 2 Cp

Run 3 Cp

CV of CP

Average of CV

STD1

12.07

14.64

14.02

13.38

4.5%

5.0%

STD2

4.02

16.35

16.08

15.17

3.9%

 

STD3

2.01

17.66

17.42

16.16

4.7%

 

STD4

1.01

19.06

18.76

17.03

6.0%

 

STD 5

0.50

20.52

20.13

18.67

4.9%

 

STD6

0.25

21.82

21.68

19.95

4.9%

 

STD7

0.13

22.59

22.07

19.79

6.9%

 

STD8

0.06

24.57

24.55

23.03

3.7%

 

STD9

0.03

24.71

24.77

22.26

6.0%

 

STD 10

0.02

25.80

25.76

23.82

4.5%

 
  1. 10-point standard synthetic v-ATPase C dsRNA was assayed in there independent runs by different analysts using the standard protocol described in Fig. 1. dsRNA amount (pg) in a 10 μL qPCR reaction and the corresponding mean (N = 3) Cp (crossing point-PCR-cycle) from each individual run are presented. The correlation between Cp and Log 2 RNA amounts are presented here. The coefficient of variation (CV) was calculated using the Cp values of each standard point