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Table 1 Effect of UNG concentration and incubation temperature 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

   0.5 U UNG 1.0 U UNG
   15°C 20°C 25°C 15°C 20°C 25°C
Analyte (dil.) Controlc CT incr. CT incr. CT incr. CT incr. CT incr. CT incr.
RNA (undil.)b 21.21 21.77 0.56 21.38 0.17 21.81 0.60 22.04 0.83 21.97 0.76 23.34 2.13
RNA (1:10) 24.91 25.11 0.20 25.29 0.38 25.44 0.53 25.10 0.19 26.23 1.32 26.69 1.78
RNA (1:100) 28.32 28.05 -0.27 28.41 0.09 29.23 0.91 28.47 0.15 29.73 1.41 30.13 1.81
RNA (1:1000) 31.16 31.78 0.62 31.59 0.43 32.15 0.99 32.70 1.54 32.70 1.54 33.19 2.03
DNA (1:107)b 21.20 24.22 3.02 26.04 4.84 27.54 6.34 24.67 3.47 28.09 6.89 30.45 9.25
DNA (1:108) 23.91 28.01 4.10 29.20 5.29 31.46 7.55 28.06 4.15 32.30 8.39 34.98 11.07
DNA (1:109) 27.09 31.71 4.62 32.89 5.80 33.50 6.41 32.00 4.91 36.02 8.93 No CTe -
DNA (1:1010) 30.48 34.43 3.95 36.00 5.52 36.24 5.76 36.78 6.30 39.03 8.55 No CT -
Mean CT increase for RNA 0.28   0.27   0.76d   0.68   1.26d   1.94d
Mean CT increase for DNA 3.92   5.36   6.52   4.71   8.19   10.16
  1. a Incubations were performed before RT-PCR with the indicated concentrations of UNG per 25 μl reaction at the indicated temperatures for 10 min.
  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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