Skip to main content

Table 2 Effect of UNG concentration on DNA degradation and RNA detectiona

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

  

0.1 U UNG

0.25 U UNG

0.5 U UNG

1.0 U UNG

Analyte (dil.)

Controlc

CT

incr.

CT

incr.

CT

incr.

CT

incr.

RNA (undil.)b

21.26

22.30

1.04

22.79

1.53

23.09

1.83

23.86

2.60

RNA (1:10)

26.25

26.86

0.61

27.57

1.32

27.95

1.70

27.31

1.06

RNA (1:100)

31.99

32.52

0.53

33.15

1.16

33.47

1.48

34.04

2.05

RNA (1:1000)

35.08

35.25

0.17

36.73

1.65

36.86

1.78

No Ct

-

DNA (1:107)b

21.53

26.56

5.03

35.04

13.51

No Ct

-

No Ct

-

DNA (1:108)

24.53

30.24

5.71

38.34

13.81

No Ct

-

No Ct

-

DNA (1:109)

28.01

33.38

5.37

No Cte

-

No Ct

-

No Ct

-

DNA (1:1010)

30.82

36.44

5.62

No Ct

-

No Ct

-

No Ct

-

Mean CT increase for RNA

0.59d

 

1.42d

 

1.70d

 

1.90d

Mean CT increase for DNA

5.43

 

13.66

 

N/A

 

N/A

  1. a Incubation was performed before RT-PCR at 30°C for 30 min at the indicated enzyme concentrations
  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