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Table 2 Detection yield, copy number and cycle threshold obtained in qPCR for the RT systems Sensiscript, Omniscript, and SuperScript II.

From: Detection limits of several commercial reverse transcriptase enzymes: impact on the low- and high-abundance transcript levels assessed by quantitative RT-PCR

Background RNA (ng)

 

0

10

25

50

100

1000

2000

RT systemsa

Datab

       

Sensiscript

        
 

DY (%)

0.62

1.36

7.69

14.57

16.64

13.61

10.63

Low copy

copy num. ± SD

2 ± 2

3 ± 4

20 ± 18

37 ± 11

43 ± 21

35 ± 19

27 ± 24

 

(Ct ± SD)

(39.52 ± 0.68)

(39.28 ± 0.71)

(37.19 ± 2.45)

(35.35 ± 0.43)

(35.34 ± 0.72)

(35.61 ± 0.93)

(36.87 ± 2.71)

High copy

DY (%)

3.92

25.61

29.79

39.26

38.77

48.33

50.41

 

copy num. ± SD

10 068 ± 8880

65 700 ± 9128

76 433 ± 833

100 733 ± 18 034

99 467 ± 9 089

124 000 ± 21 071

129 333 ± 34 020

 

(Ct ± SD)

(30.6 ± 7.14)

(24.24 ± 0.21)

(24.02 ± 0.02)

(23.62 ± 0.28)

(23.64 ± 0.14)

(23.29 ± 0.29)

(23.67 ± 0.14)

GNPDA

copy num. ± SD

0 ± 0

73 ± 25

236 ± 37

691 ± 39

2 150 ± 806

26 395 ± 1 404

42 488 ± 6 724

 

(Ct ± SD)

(40 ± 0)

(32 ± 0.62)

(30.03 ± 0.3)

(28.21 ± 0.09)

(26.42 ± 0.58)

(22.22 ± 0.09)

(21.45 ± 0.26)

Omniscript

        

Low copy

DY & copy num.

N/A

N/A

N/A

N/A

N/A

N/A

N/A

 

DY (%)

0.00

0.00

0.00

0.24

2.16

38.46

38.56

High copy

copy num. ± SD

0 ± 0

0 ± 0

9 ± 15

623 ± 368

5 548 ± 4 152

98 664 ± 5 062

98 927 ± 11 165

 

(Ct ± SD)

(40 ± 0)

(40 ± 0)

(38.41 ± 2.75)

(30.73 ± 1.59)

(27.53 ± 2.1)

(22.44 ± 0.08)

(22.44 ± 0.17)

GNPDA

copy num. ± %

(Ct ± SD)

primer dimer

primer dimer

primer dimer

primer dimer

primer dimer

primer dimer

primer dimer

SuperScript II

        
 

DY (%)

19.80

49.43

70.14

66.30

81.33

102.44

97.38

Low copy

copy num. ± SD

51 ± 44

127 ± 39

180 ± 18

170 ± 49

209 ± 12

263 ± 40

250 ± 106

 

(Ct ± SD)

(34.05 ± 0.04)

(33.41 ± 0.51)

(32.72 ± 0.19)

(32.82 ± 0.42)

(32.47 ± 0.09)

(32.15 ± 0.21)

(32.32 ± 0.7)

 

DY (%)

13.30

22.33

16.69

18.86

27.31

31.33

36.81

High copy

copy num. ± SD

34 127 ± 4 284

57 277 ± 5 106

42 814 ± 26,175

48 389 ± 24 717

70 062 ± 8 760

80 366 ± 19 132

94 447 ± 10 237

 

(Ct ± SD)

(23.52 ± 0.19)

(22.75 ± 0.14)

(23.48 ± 1.19)

(23.18 ± 0.98)

(22.45 ± 0.19)

(22.28 ± 0.4)

(22 ± 0.16)

GNPDA

copy num. ± SD

5 ± 9

102 ± 48

351 ± 201

1 122 ± 608

2 401 ± 752

19 088 ± 3 540

22 758 ± 1 815

 

(Ct ± SD)

(38.79 ± 2.1)

(30.84 ± 1.17)

(28.6 ± 1.12)

(26.43 ± 0.91)

(24.95 ± 0.57)

(21.2 ± 0.35)

(20.86 ± 0.14)

  1. aRT systems tested were Sensiscript. Omniscript. SuperScript II. SuperScript III. and PowerScript; low copy. RT samples spiked with 2.6 × 103 EGFP mRNA copies; high copy. RT samples spiked with 2.6 × 106 EGFP mRNA copies.
  2. bDY (detection yield) was calculated using the equation described in Results and the copy number (num.) was derived from the calibration curve for a given cycle threshold value (CT) [see Additional file 2];SD. standard deviation.
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BMC Molecular Biology

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