Relative quantification of mRNA: comparison of methods currently used for real-time PCR data analysis

BMC Molecular Biology

Table 2 Pearson's correlation coefficients obtained from the linear regression plotting R₀ values against diluting factors of the total RNA (RT-PCR template). The R₀ values were obtained by transformation of fluorescence data using the following methods for real-time PCR data analysis: St cur, relative standard curve; Comp, comparative Ct; SCF, sigmoid curve-fitting; DART ind E, DART-PCR with individual E values; DART av E, DART-PCR with average E values; Liu&S ind E, Liu & Saint-exp with individual E values, Liu&S av E, Liu & Saint-exp with average E values; LinReg ind E, LinRegPCR (using individual E values); LinReg-Ct av E, LinRegPCR combined with Ct (using average E values).

Gene/Method	St cur	Comp	SCF	DART ind E	DART av E	Liu&S ind E	Liu&S av E	LinReg ind E	LinReg-Ct av E
IL-1β	0.9993	0.9996	0.9960	0.9924	0.9997	0.9697	0.9994	0.9113	0.9994
IL-6	0.9998	0.9998	0.9951	0.9745	0.9993	0.9510	0.9996	0.9391	0.9997
TNF-α	0.9996	0.9998	0.9987	0.9910	0.9989	0.9622	0.9996	0.9835	0.9996
GM-CSF	0.9980	0.9980	0.9803	0.9620	0.9985	0.9277	0.9975	0.9426	0.9980
ACTB	0.9991	0.9992	0.9973	0.9435	0.9972	0.9915	0.9998	0.9828	0.9984
SDHA	0.9998	0.9999	0.9999	0.9799	0.9999	0.9872	0.9999	0.9902	0.9999
HPRT	0.9996	0.9995	0.9997	0.9699	0.9990	0.9662	0.9994	0.9545	0.9997
Average	0.9991	0.9994	0.9953	0.9733	0.9990	0.9651	0.9993	0.9577	0.9992

ISSN: 1471-2199