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Table 2 Housekeeping gene validation in the pMCAO model

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

  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)ΔCT 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