Primer design¶

In some of these videos, the interface might be slightly different, but the functionality is basically the same.

Parameters used to calculate melting temperature for primer design

You can set the parameters used to calculate melting temperature in the Settings tab.

Primer design for homologous recombination¶

Primer design for CRISPR-HDR¶

You can design primers to add homology arms when amplifying the donor sequence, the same way you would do for homologous recombination (see above).

Primer design for Gibson Assembly¶

Designing primers to circularize a single fragment using Gibson Assembly¶

When you are in the PCR form, and you select Design primers, then Gibson Assembly, don't select any other input sequence, and click Design primers. Don't forget to select Circular Assembly in the Other settings field of the primer design form.

Primer design for In Vivo Assembly¶

The process is similar to Gibson Assembly, simply select "In Vivo Assembly" in the primer design form.

Primer design for In-Fusion Cloning¶

The process is similar to Gibson Assembly, simply select "In-Fusion Cloning" in the primer design form.

Primer design for Overlap Extension PCR¶

The process is similar to Gibson Assembly, use the Gibson Assembly primer design form, then change the method to "Overlap Extension PCR".

Primer design for Golden Gate Assembly¶

Coming soon