In this mode the user must define either a Forward primer
, a Reverse primer region
, or both. These are defined
by making a selection on the sequence and right-clicking the
selection. It is also possible to define a Region to amplify
in which case a forward- and a reverse primer region are
automatically placed so as to ensure that the designated region will
be included in the PCR fragment. If areas are known where primers
must not bind (e.g. repeat rich areas), one or more No primers
regions can be defined.
If two regions are defined, it is required that at least a part of the
Forward primer region is located upstream of the
Reverse primer region.
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.