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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
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