Constructed standard curves for the determination of the PCR amplification efficiencies. Serial diluted 1:5 cDNA dilutions from the #1 MHS muscle biopsy. Each standard curve represents the pooled results of four individual experiments. Amplifications in each individual experiment were conducted in triplicate. Housekeeping gene; HRPT (triangles), wild type RYR 1; AS1 (diamonds) and mutant RYR 1; AS2 (squares). The error bars show the standard deviation. The regression coefficient was greater than 0.99 in all cases.