TABLE 2.

Effects of diluent and UNG on T_m values of HSV-2-specific probes

Diluent	Dilution	Copy no.a	Cp	Genotype	Tm	ΔTm
H2O (0.5 U UNG)	10−1	107	32.63	HSV-2	66.33	−0.20
	10−2	5	37.54	UKN	59.55	−6.98c
	10−3	2	39.44	UKN	56.46	−10.07c
H2O (no UNG)	10−1	93	32.81	HSV-2	66.15	−0.38
	10−2	18	35.42	UKN	61.07	−5.46c
	10−3	2	39.36	UKN	56.38	−10.15c
10 mM Tris, pH 8.0 (0.5 U UNG)	10−1	95	32.82	HSV-2	66.82	0.29
	10−2	8	36.59	UKN	60.70	−5.83c
	10−3	1	>40.00	UKN	59.25	−7.28c
10 mM Tris, pH 8.0 (no UNG)	10−1	75	33.15	HSV-2	66.63	0.10
	10−2	13	35.92	UKN	61.24	−5.29c
	10−3	1	>40.00	UKN	54.91	−11.62c
EB,b pH 8.0 (0.5 U UNG)	10−1	93	32.85	HSV-2	66.63	0.10
	10−2	8	35.40	UKN	60.98	−5.55c
	10−3	1	>40.00	UKN	54.91	−11.62c
EB, pH 8.0 (no UNG)	10−1	77	32.55	HSV-2	66.83	0.30
	10−2	3	35.79	UKN	60.87	−5.66c
	10−3	2	39.22	UKN	59.65	−6.88c

^aThe copy number was estimated in relation to the copy number on a standard curve generated with plasmid-borne controls.

^bEB, elution buffer provided with the MagNA Pure LightCycler total nucleic acid kit.

^cCompared to the T_m values for the HSV-1- or HSV-2-positive control, the ΔT_m values were above the acceptable variation (±2.5°C) recommended by the manufacturer.