Table 1.

An example of four-parameter sigmoidal equation modeling and result of fluorescence normalization

Cycle Numberxa	D2 sample FAM (Raw Rx)b	D2 Normalized Rx . (Raw Rx -/Raw Y0-1)c
23.58	353.48	0.056045583
24.58	355.38	0.061721963
25.58f	367.68	0.098469051
26.58	380.74	0.13748669
27.58	409.2	0.222512879
28.56	429.79	0.284026907
29.61	465.57	0.390922095
30.56	504.15	0.506182473
31.63	535.18	0.598886713
32.63	576.31	0.72176539
33.63	595.56	0.779276076
34.63	639.21	0.909683425
35.63	669.3	0.99957935
Parameter	Sigmoid function fitted for raw fluorescenced	Sigmoid function fitted for normalized fluorescencee
a	525.5884	1.5702
b	3.2674	3.2674
X0	33.4481	33.4481
Y0	334.7204	0
R	0.998805	0.99880514

^a amplification cycle number

^b Raw FAM fluorescence data

^cNormalized fluorescence = Raw Rx/Raw y_o-1

^d 4 parameter sigmoid function fitted for raw fluorescence

^e 4 parameter sigmoid function fitted for normalized fluorescence

^f bold text cycles are selected PCR exponential phase for allele ratio quantification