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Table 2 Relative allele frequencies using engineered plasmid constructs

From: Allele-specific differences in ryanodine receptor 1 mRNA expression levels may contribute to phenotypic variability in malignant hyperthermia

Ratio (WT:MT)

Ct values

Percentage (%)

  

Mean (n = 3)

Stdev.

ΔCt

bΔΔCt

Theor.

Obs.

4:1

AS1

18.26

0.09

-3.35

-2.10

80.00

81.09

 

AS2

21.61

0.09

3.35

2.10

20.00

18.91

3:1

AS1

18.50

0.06

-2.72

-1.47

75.00

73.43

 

AS2

21.21

0.08

2.72

1.47

25.00

26.57

1:2

AS1

19.55

0.07

-0.21

1.04

33.33

32.67

 

AS2

19.76

0.04

0.21

-1.04

66.67

67.33

1:1

AS1

18.95

0.21

a-1.25

0.00

-

-

 

AS2

20.20

0.08

a1.25

0.00

-

-

  1. Stdev. = Standard deviation, Theor. = theoretical values, Obs. = experimentally observes values. a The ΔCt value for the 1:1 ratio should be equal to zero. Any deviation indicates differences in amplification efficiencies or DNA concentrations of the added plasmid constructs. Therefore the 1:1 ΔCt values were subtracted from the ΔCt values leading to b ΔΔCt. This value was used in Equation 1 to calculate the relative allele frequencies.