Table 2. Optimal ranges of hybridization temperature, NaCl and denaturant concentration predicted through the RSM models for the tested bacteria.

The predicted and experimental fluorescence values are also shown. Optimum ranges for each parameter have been stablished assuming a fluorescence intensity of at least 85% of the maximum value.

Denaturant	Bacteria	Hybridization conditions			Predicted fluorescence (a.u.)	Obtained fluorescence (a.u.)*
		Temperature (°C)	[NaCl] (M)	[Denaturant] (M; % v/v)
Formamide	E. coli	51.4–76.1	0.03–1.99	1.18–4.40	296.13–348.87	369.14 ±32.62
	P. aeruginosa	56.4–74.2	2.11–4.92	4.27–12.80	322.97–379.97	225.30 ± 14.58
	C. freundii	50.0–79.0	2.25–5.00	9.00–17.69	113.76–133.83	141.26 ± 38.86
	E. faecalis	50.0–69.3	2.66–5.00	22.00–32.81	288.31–339.19	131.15 ± 4.31
	S. epidermidis	62.0–82.0	2.00–5.00	3.00–13.00	47.51–55.89	296.62 ± 46.55
Ethylene carbonate	E. coli	55.5–77.9	2.00–5.00	3.13–12.42	278.02–327.08	346.03 ± 19.93
	P. aeruginosa	53.8–78.1	2.03–3.42	4.02–12.71	196.01–230.61	291.57 ± 14.15
	C. freundii	57.9–79.0	2.00–4.89	3.76–13.00	266.56–313.59	353.14 ± 12.17
	E. faecalis	46.0–63.0	2.00–5.00	1.00–4.00	174.27–205.03	126.34 ± 20.76
	S. epidermidis	62.0–82.0	2.90–5.00	0.00–2.00	106.68–125.50	254.37 ± 17.45
Urea	E. coli	50.1–60.0	2.41–5.01	1.41–4.09	189.217–236.52	253.87 ± 13.73
	P. aeruginosa	47.1–58.1	2.45–5.01	0.67–2.37	395.90–465.75	324.35 ± 18.26
	C. freundii	50.2–63.3	2.42–4.57	0.63–2.08	183.51–215.89	225.82 ± 39.74
	E. faecalis	50.1–79.9	2.59–5.01	1.41–4.09	345.03–405.92	205.58 ± 17.75
	S. epidermidis	64.2–82.9	3.37–5.01	0.61–1.84	188.04–221.22	314.17 ± 21.52

*The obtained fluorescence was evaluated using the optimum hybridization temperature, denaturant and salt concentration predicted through the RSM models for each bacterium.