Real time relative quantitation of Wnt 10b and Wnt 11 expression in DD vs. control. Real time relative quantitation of PCR amplicons was performed on an ABI Prism 7700 using the comparative Ct method. UPPER PANELS: Determination of relative amplification efficiencies for Wnt10b vs. β-actin and Wnt11 vs. β-actin. As shown, target and endogenous control gene products amplified with similar efficiency. The Δ CT value (CTtarget/CTendogenous control) was determined to be 0.1696 and 0.1619 for Wnt 10b/β-actin and Wnt 11/β-actin respectively. LOWER PANEL: Real time relative PCR quantitation for Wnt10b and Wnt11 mRNAs performed on a separate subset of 7 DD and 6 PF control samples derived from surgical resection. Triplicate reactions of each dilution were performed (50 μl samples) in a 96-well plate format using SDS instrumentation (Applied Biosystems) for 45 cycles. Target and endogenous control reaction were run in separate wells in triplicate at each concentration. Relative Quantitation of Wnt 10b and Wnt 11 mRNA levels in normal palmar fascia (mean Wnt 10b = 20.74 ± 8.81, Wnt 11 = 40.89 ± 12.86) were significantly (*) different to DD (Wnt 10b = 4.83 ± 2.21; P = 0.038, Wnt 11 = 7.56 ± 4.50; P = 0.014, Mann Whitney Test). Data is shown as Mean ± SEM.
O'Gorman et al. Journal of Negative Results in BioMedicine 2006 5:13 doi:10.1186/1477-5751-5-13