Acquisition of data at multiple gains within a single thermal melt experiment using the Rotor-Gene Q instrument

Journal of Analytical Science and Technology

Table 1 Δ T _m of protein for ten compounds (C1 to C10)

Concentration of compounds C1 to C10 (mM)		The values of Δ T _m at different gains used in the same experimental run (°C)					Average Δ T _m (°C)
Concentration of compounds C1 to C10 (mM)		Gain −2	Gain 0	Gain 2	Gain 4	Gain 6	Average Δ T _m (°C)
0.5		0.83	0.93	0.93	0.63	-	0.82 ± 0.14
2	C1	1.13	1.63	1.43	-	-	1.39 ± 0.25
5		1.63	2.13		-	-	1.87 ± 0.35
0.5		0.83	1.33	1.13	0.93	-	1.05 ± 0.22
2	C2	1.13	1.63	1.63	1.33	-	1.42 ± 0.24
5		1.13	1.63	1.83		-	1.52 ± 0.36
0.5		0.33	0.83	0.63	0.63	-	0.60 ± 0.21
2	C3	−0.08	0.33	0.13	0.13	-	0.12 ± 0.16
5		0.43	0.33	0.43	0.13	-	0.32 ± 0.14
0.5		0.63	1.13	1.33	0.93	-	1.00 ± 0.30
2	C4	2.93	3.63	3.43	3.13	-	3.27 ± 0.31
5		−0.08	0.43	0.63		-	0.32 ± 0.36
0.5		0.43	0.63	0.63	0.43	-	0.52 ± 0.12
2	C5	−0.18	0.13	0.13	−0.18	-	−0.03 ± 0.17
5		0.33	0.63	0.83	-	-	0.59 ± 0.25
0.5		0.63	0.83	0.63	0.33	-	0.60 ± 0.21
2	C6	0.63	0.83	0.63	0.33	0.33	0.55 ± 0.22
5		0.33	0.33	0.13	−0.08	−0.18	0.11 ± 0.23
0.5		0.43	1.43	0.93	-	-	0.93 ± 0.05
2	C7	−0.18	0.13	-	-	-	−0.03 ± 0.21
5		-	-	-	-	-	-
0.5		1.13	1.43	1.13	0.83	0.63	1.03 ± 0.31
2	C8	1.13	1.63	1.63	1.33	-	1.43 ± 0.24
5		2.43	2.83	3.13		-	2.79 ± 0.35
0.5		-	0.33	0.13	−0.38	−0.38	−0.08 ± 0.36
2	C9	-	1.33	1.13	0.93	0.83	1.05 ± 0.22
5		2.63	3.63	3.63	3.13	3.13	3.23 ± 0.42
0.5		0.43	0.83	0.63	0.43	-	0.58 ± 0.19
2	C10	1.13	1.63	1.43	1.13	-	1.33 ± 0.24
5		1.83	2.33	2.33	1.93	-	2.10 ± 0.26

Hyphen indicates samples that did not show fluorescence melting signal above threshold noise level or resulted in signal saturation (data loss) due to high fluorescence intensity. The ΔT _m of protein was measured for the ten compounds C1 to C10 at three concentrations (0.5, 2, and 5 mM) at five levels of gain set within a single experimental run. T _m of the protein (T _m ^ref) = 58.4 ± 0.2°C.