Real time quantitative PCR of individual microRNA.

<p>From the microRNA (miRNA) array analysis 16 miRNAs were identified as having significant differential expression (p<0.05) and selected for further validation with real time quantitative PCR (RT-qPCR). Threshold cycle (Ct) values were normalised using the average Ct of small nucleolar RNAs (snoRNAs) and to determine miRNA relative expression 2ΔΔCT was used and calculated as follows ΔCT (miRNA Ct—averaged endogenous control Ct) and fold-change to reference sample of normal liver (a non steatotic donation after brain stem death liver). Due to the range and magnitude of relative miRNA expression, data was log transformed and presented as mean +/- SE. The relative expression for a given miRNA in the three donation after cardiac death (DCD) liver groups of primary non function (PNF) retransplanted within a week (n = 7), good functional outcome (n = 7) peak aspartate transaminase (AST) ≤ 1000 IU/L and early graft dysfunction (EGD) (n = 9) peak AST ≥ 2500 IU/L is represented graphically. The observed differences in the relative expression of a given miRNA species on RT-qPCR between the three DCD liver groups was only found to be significant for miR-22 (p = 0.049).