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Table 3 Effect of incubation temperature on DNA degradation and RNA detection in the presence of UNGa

From: Quantitative assessment of the effect of uracil-DNA glycosylase on amplicon DNA degradation and RNA amplification in reverse transcription-PCR

  

25°C

30°C

35°C

Analyte (dil.)

Controlc

CT

incr.

CT

incr.

CT

incr.

RNA (undil.)b

21.22

22.40

1.18

23.17

1.95

23.79

2.57

RNA (1:10)

26.15

26.93

0.78

27.54

1.39

28.25

2.10

RNA (1:100)

31.37

32.51

1.14

32.05

0.68

33.79

2.42

RNA (1:1000)

33.62

35.46

1.84

35.24

1.62

37.43

3.81

DNA (1:107)b

21.48

32.11

10.63

37.51

16.03

38.57

17.09

DNA (1:108)

24.89

34.33

9.44

No Cte

-

No Ct

-

DNA (1:109)

28.22

36.08

7.86

No Ct

-

No Ct

-

DNA (1:1010)

31.75

38.48

6.73

No Ct

-

No Ct

-

Mean CT increase for RNA

1.24d

 

1.41d

 

2.73d

Mean CT increase for DNA

8.67

 

16.03

 

17.09

  1. a The UNG concentration was 0.25 units per 25 μl reaction and incubation time before RT-PCR was 30 min at the indicated temperatures.
  2. b The undiluted RNA sample and the 1:107 dilution of the DNA sample contained 250,000 copies of RNA or DNA, respectively
  3. c Control reactions did not contain UNG and were not incubated prior to RT-PCR
  4. d P < 0.05 by paired t-test compared to control values
  5. e Indicates that amplification was not detected through 40 cycles in each of three replicate reactions