Ulrich Krauss and
Thorsten Eggert (2005) "insilico.mutagenesis:
a primer selection
tool designed for sequence scanning applications used in directed evolution
experiments." BioTechniques 39(5): p679-682
This online tool allows the prediction of large sets of oligonucleotide primers used in saturation mutagensis and scanning mutagenesis experiments.
Read the article mentioned above for more information.
The program generates the so-called mutagenesis primers to perform subsequent single amino acid exchanges within a defined region. Oligo's are designed in accordance with the MegaPrimer PCR approach as described by Barettino et al (1994). For further information see the figure below.
proceed to insilico.mutagenesis
The principle of the MegaPrimer PCR technique:
The online tool: insilico.mutagenesis will generate the in the figure highlighted mutagenesis primer which is used in the first PCR reaction of the megaprimer method.