Linearity
Linearity was measured according to CLSI Guideline EP6-A “Evaluation of the linearity of quantitative measurement procedures: a statistical approach; approved guideline”.
Plasmids carrying wild type and mutant sequences were mixed in proportions to give the following levels of mutation: 0, 12.5, 25, 37.5, and 50%. Four replicates of the mixes were placed in a random pattern on a plate and analyzed. The results for the mutation GGT->TGT in codon 12 are shown (figure " Linearity of GGT->TGT mutation").
Precision
Determination of the linearity of the mutation GGT->TGT in codon 12 was repeated by 3 operators on 3 separate days using different combinations of PyroMark Q24 instruments and reagents.
The values for the intermediate precision (SD) were 0.6–2.0 % units in the measured range of 0–50% mutation level.
Intermediate precision % mutated plasmid | Run 1 (Mean, SD) | Run 2 (Mean, SD) | Run 3 (Mean, SD) | Summary (Mean, SD) |
0.0 | 0.6, 0.2 | 1.7, 0.7 | 0.7, 0.2 | 1.0, 0.6 |
12.5 | 13.3, 1.5 | 16.2, 1.9 | 14.6, 3.0 | 14.7, 1.4 |
25.0 | 23.8, 1.4 | 26.8, 2.4 | 26.9, 2.9 | 25.8, 1.8 |
37.5 | 42.0, 1.9 | 41.7, 0.5 | 38.5, 2.6 | 40.7, 2.0 |
50.0 | 49.7 2.4 | 50.5, 1.8 | 49.1, 4.8 | 49.8, 0.7 |