In this mode the user can define a number of Forward primer regions and Reverse primer regions where a sequencing primer can start. These are defined by making a selection on the sequence and right-clicking the selection. If areas are known where primers must not bind (e.g. repeat rich areas), one or more No primers here regions can be defined.
No requirements are instated on the relative position of the regions defined.
After exploring the available primers (see Graphical display of primer information) and setting the desired parameter values in the Primer Parameters preference group, the Calculate button will activate the primer design algorithm.
After pressing the Calculate button a dialog will appear (see figure 19.11).
Since design of sequencing primers does not require the consideration of interactions between primer pairs, this dialog is identical to the dialog shown in Standard PCR mode when only a single primer region is chosen. See the section 19.5 for a description.