By clicking on the computation button , you can compute all active constraints for the input primer set. To select only the subset of primers fulfilling the active constraints, you can use the filtering button .
When filtering, only primers fulfilling all of the the active constraint settings are retained. For example, when the GC ratio is constrained to be between 40% and 60%, then only primers with GC ratios in this range are outputted. When computing the primer properties, the selected criteria are evaluated for all of the primers and the primers are annotated with their corresponding properties. Here, the allowed value ranges of the criteria are only used to annotate whether the primers would have fulfilled the criteria, but no filtering is performed. An application scenario for the filtering operation is the refinement of an existing set of primers, in order to obtain a more stringent set of primers.
The evaluation operation is useful to gain insights about the properties of a primer data set. For example, one might want to know the properties of all primers in a set to identify potential problems. After the evaluation has been performed, visualizations indicating the quality of the primer set can be obtained.
The presence of restriction sites in the input primers can influence the evaluation results, particularly the coverage computations. By clicking on the button, you can make sure that no restriction sites are present in the input primers.