Table 3.

Effect of incubation temperature on DNA degradation and RNA detection in the presence of UNG^a

		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

^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.

^b The undiluted RNA sample and the 1:10⁷ dilution of the DNA sample contained 250,000 copies of RNA or DNA, respectively

^c Control reactions did not contain UNG and were not incubated prior to RT-PCR

^d P < 0.05 by paired t-test compared to control values

^e Indicates that amplification was not detected through 40 cycles in each of three replicate reactions