|Stage 1||Stage 2|
|Cycles||Temperature (°C)||Time (seconds)||Cycles||Temperature (°C)||Time (seconds)|
* = Data collection step i.e. optics ON
- For some instruments it may be necessary to increase the annealing step to 50 seconds in order to achieve a robust
|Agilent Technologies / Stratagene||FAM||HEX|
- If given the option, set the Reference Dye to None.
- For some instruments it may be necessary to calibrate (colour compensate) before use to prevent cross-talk
Data analysis and interpretation
- Thresholds should be set to appropriate levels which are valid between runs.
- A Limit of Blank should be established using DNA samples that are free of Aspergillus in order to determine a
cut-off between positive and negative samples.
- For negative reactions, a viable IAC should also be reported in order to avoid false-negative results. This is normally considered to be within ± 3 standard deviations from the average.