Virology Journal

Table 1 Effect of UNG concentration and incubation temperature on DNA degradation and RNA detection^a

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.)	Control^c	C_T	incr.	C_T	incr.	C_T	incr.	C_T	incr.	C_T	incr.	C_T	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 C_T^e	-
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 C_T	-
Mean C_T increase for RNA			0.28		0.27		0.76^d		0.68		1.26^d		1.94^d
Mean C_T increase for DNA			3.92		5.36		6.52		4.71		8.19		10.16

^a Incubations were performed before RT-PCR with the indicated concentrations of UNG per 25 μl reaction at the indicated temperatures for 10 min.
^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

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ISSN: 1743-422X

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