Validation of housekeeping genes for quantitative real-time PCR in in-vivo and in-vitro models of cerebral ischaemia

Table 2 Housekeeping gene validation in the pMCAO model

	Harrison DC 2000	Medhurst AD 2000	Meldgaard M 2006	Tian YF 2007	Gubern C
Focal ischaemia model	pMCAO intraluminal filament	pMCAO intraluminal filament	pMCAO electrocautery	pMCAO intraluminal filament	pMCAO electrocautery
Animal	Male Sprague-Dawley rats	Male Sprague-Dawley rats	Male SJL mice	Male Sprague-Dawley rats	Male Fisher rats
Collected sample	Whole infarcted cortex	Whole infarcted cortex	Whole infarcted hemisphere	Ischaemic penumbra cortex	Infarcted and peri-infarcted cortex region
Time points	Naive, 3, 6, 12 and 24 h (pMCAO and sham)	Contralateral hemisphere, 6 and 24 h	Sham, 30 min, 1, 2, 4, 6, 12, 24 and 48 h, 5 and 10d	Sham, 2 h, 24 h, 3, 7 and 21d	Sham, 30 min, 2, 6, 24, 48 and 72 h
Real-Time PCR	Custom TaqMan assays	Custom TaqMan assays	Custom TaqMan assays	SyberGreen	Inventoried TaqMan assays
Method	Standard curve	Standard curve	%mRNA = 100 × (1+E)Δ^CT	2^-ΔC'T	2^-ΔC'T
Statistical analyses	ANOVA	Student's unpaired t-test	Kruskal-Wallis + Dunn's	ANOVA + Tukey	ANOVA + Dunnett's
Suitable controls	Ppia & Gapdh	Ppia & Gapdh	Hprt & Gapdh	Rpl13a & Ppia	Hprt, Sdha & Ywhaz
Unsuitable controls	β-actin			Gapdh, β-actin	Oaz1, Gapdh, Ppia, Rpl13a, B2m, Gusb, 18SrRNA

ISSN: 1471-2199