How can I create a primer?
To create a primer, click Primers → Add Primer... , then copy and paste a sequence.
Alternatively, you can begin by selecting the desired binding site. If you click the mouse and drag to select, the melting temperature of a corresponding primer will be shown.
If the primer will be made from a selected binding site, specify whether the Top Strand or Bottom Strand of the selection should be used.
If desired, type the primer name.
If desired, type a description. The Description tab is open by default.
If desired, modify the primer to add a 5’ extension or introduce a mutation. There are two options.
One option is to manually edit the primer sequence in the text box.
The other option is to click in the primer sequence at the location of the change, and then click the Insertions tab. Use the dialog controls to add a desired codon, restriction site, or peptide coding sequence.
Note the number of binding sites and the calculated melting temperature. To see a summary of the melting temperature calculation methods, click the blue text at the lower right corner of the dialog.
Click Add Primer to Template to see the new primer.