Table 3.

Comparison of the sensitivity of TaqMan and SYBR Green real time PCR assays. All reactions were performed with the same plasmid standard, except in cases where the appropriate plasmid standard encompassing both primer sets was not available, and a cDNA sample was used instead. The same samples were used to compare the TaqMan assays in this study with either previously published TaqMan assays or previously published SYBR Green assays for an individual gene. Individual lines indicate different experiments.

Gene target	Sensitivity of real time reactiona						References for other published studies
	TaqMan primer/probes from this study		TaqMan primer/probes from other published studies		SYBR Green primers from other published studies
	TaqMan assay		TaqMan Assay		SYBR Green assay
	Copy number/RNA amount	Average Ct ± std dev	Copy number/RNA amount	Average Ct ± std dev	Copy number/RNA amount	Average Ct ± std dev
IL1α	20.8b	34.8 ± 0.1			n.a.	n.a.
IL1β	21.9b	33.8 ± 0.3			21.9b	35.3 ± 0.2	Fang et al. (2010) and Rowe et al. (2010)
IL2	26.3b	34.6 ± 0.3			26.3b	29.9 ± 0.8	Svitek and von Messling (2007)
IL4	26.6b	34.7 ± 0.5			26.6b	35.3 ± 0.4	Maines et al. (2012)
IL4	26.6b	34.7 ± 0.5			26.6b	33.2 ± 0.1.	Svitek and von Messling (2007)
IL6	28.9b	36.8 ± 0.3			28.9b	33.6 ± 0.9	Svitek and von Messling (2007)
IL6	0.1 ng	38.2 ± 0.6	0.1 ng	38.7 ± 0.7			Nakata et al. (2009)
IL8	26.9b	37.1 ± 0.6			26.9b	34.0 ± 0.4	Maines et al. (2012)
IL10	27.1b	35.0 ± 0.3			27.1b	33.9 ± 0.9	Svitek and von Messling (2007)
IL10	27.1b	33.3 ± 0.5	27.1b	34.6 ± 0.5			Nakata et al. (2009)
IL12p40	29.6b	33.2 ± 0.4			n.a.d	n.a.d
IL12p40d	1 ng	33.9 ± 0.4			1 ng	34.6 ± 1.2	Svitek and von Messling (2007)
IL17	28.2b	33.9 ± 0.4			n.a.	n.a.
Granzyme A	22.8b	35.3 ± 0.7			n.a.	n.a.
MCP1	29.0b	34.9 ± 0.2			n.a.d	n.a.d
MCP1d	0.1 ng	33.3 ± 0.3			1 ng	34 ± 0.9	Rowe et al. (2010)
TNFα	29.1b	34.1 ± 0.6			29.1b	31.1 ± 0.6	Svitek and von Messling (2007)
TNFαd	0.1 ng	33.7 ± 0.6			0.1 ng	34.5 ± 0.7	Fang et al. (2010) and Rowe et al. (2010)
TNFα d	0.1 ng	34.6 ± 0.5	0.1 ng	35.0 ± 0.6			Nakata et al. (2009)
IFNα	22.9b	34.7 ± 0.8			22.9b	34.5 ± 0.8	Svitek and von Messling (2007)
IFNβ	22.5b	35.7 ± 0.7			22.5b	31.6 ± 0.4	Maines et al. (2012)
IFNγ	26.3b	35.1 ± 0.5			26.3b	33.6 ± 1.1	Svitek and von Messling (2007)
IFNγ	26.3b	34.8 ± 0.2	26.3b	35.9 ± 0.1			Nakata et al. (2009)
Matrix	23.3b	37.2 ± 0.8			n.a.	n.a.
ATF4	28.9b	33.9 ± 0.5			n.a.	n.a.
HPRT	29.1b	35.9 ± 0.3			n.a.	n.a.
GAPDH	<10c	37.6 ± 0.9			2.8c	37.8 ± 1	Svitek and von Messling (2007)
GAPDHd	0.3 ng	33.4 ± 1.5	0.03 ng	31.7 ± 0.6			Nakata et al. (2009)
L32	29.3b	36.6 ± 0.7			n.a.	n.a.

n.a.: not applicable.

^aSensitivity refers to the minimum number of copies detected after serial ten-fold dilutions of DNA plasmid, in all triplicates. All three replicates must have a detectable Ct value for inclusion.

^bCopy number is equivalent to 0.0001 pg DNA plasmid standard.

^cCopy number is equivalent to 0.00001 pg DNA plasmid standard.

^dPlasmid did not overlap with previously published primer sequence.