Evaluation of reference genes for real-time quantitative PCR studies in Candida glabrata following azole treatment

Table 2 Stability of RNA transcription of reference genes in fluconazole-treated C. glabrata as determined by the 2 ^-ΔΔCT method

	RDN18	RDN25	RDN5.8	UBC13	PGK1	UBC7	GAPDH	UBC4	TUB1	EF1α	PPIA	RPL2A	RPL10	RPL13A	ACT1	SDHA
C _T Change	0.34	0.39	0.46	1.34	1.56	1.60	2.17	2.41	2.60	2.89	3.13	3.67	4.12	4.17	4.45	5.03
ΔΔC _T (t)	0.12	0.07	0	-0.87	-1.10	-1.13	-1.71	-1.95	-2.13	-2.43	-2.66	-3.21	-3.65	-3.71	-3.98	-4.57
2 ^-ΔΔCT (t)	0.92^#	0.95^#	1.00	1.83*	2.14*	2.19*	3.28*	3.86*	4.39*	5.38*	6.32*	9.23*	12.58*	13.06*	15.82*	23.75*
Ranking	1	2	3	4	5	6	7	8	9	10	11	12	13	14	15	16

C_T Change = C_T(ut) − C_T(t); ut, untreated; t, fluconazole-treated.
ΔC_T = C_T_reference − C_T_RDN5.8; ΔC_T of each reference gene was calculated using RDN5.8 as the internal control.
ΔΔC_T(t) = ΔC_T(t) − ΔC_T(ut).
2^-ΔΔCT(t) for fluconazole-treated cells indicates fold change in RNA transcription of a reference gene normalized to RDN5.8, as compared with untreated cells.
The stability ranking is based on the values of C_T Change, ΔΔC_T(t), and 2^-ΔΔCT(t) of the reference genes in fluconazole-treated C. glabrata cells.
^#P > 0.05 and *P <; 0.05 for the azole-treated group vs. the untreated group after normalizing to RDN5.8.

ISSN: 1471-2199