Site-Directed Mutagenesis Primer Designer (Beta)

Design primers for single or multiple nucleotide substitutions in one final sequence.

Mutagenesis primer inputs

Template DNA

Nucleotide changes

Use 1-based positions in the original sequence.

Primer settings

Editable design limit; primers over 60 nt need synthesis review.

Reaction settings

References

  1. NEB: back-to-back mutagenesis primer design and multiple-site overlap mutagenesis.
  2. Jin, Kim and Ignea (2026): PrimerWeaver, including reported overlap-mutagenesis sequences.
  3. Binding-region Tm: SantaLucia (1998) and Owczarzy et al. (2008). Calculations use Mol Biology Tools code.