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Table 4 Effect of incubation time 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

   10 min 20 min 30 min
Analyte (dil.) Controlc CT incr. CT incr. CT incr.
RNA (undil.)b 21.07 21.79 0.72 22.69 1.62 22.74 1.67
RNA (1:10) 25.80 25.96 0.16 26.83 1.03 27.51 1.71
RNA (1:100) 31.28 31.74 0.46 32.15 0.87 32.78 1.50
RNA (1:1000) 34.81 36.16 1.35 36.01 1.20 35.6 0.79
DNA (1:107)b 21.46 28.19 6.73 32.71 11.25 35.93 14.47
DNA (1:108) 24.73 31.22 6.49 36.52 11.79 No Ct -
DNA (1:109) 28.08 34.60 6.52 No Ct - No Ct -
DNA (1:1010) 31.14 No Cte - No Ct - No Ct -
Mean CT increase for RNA 0.67   1.18d   1.42d
Mean CT increase for DNA 6.58   11.52   14.47
  1. a The UNG concentration was 0.25 units per 25 μl reaction and incubation before RT-PCR was performed at 30°C for the indicated times.
  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
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