Open Accessinsilico.mutagenesis: a primer selection tool designed for sequence scanning applications used in directed evolution experiments
Author(s) -
Ulrich Krauß,
Thorsten Eggert
Publication year - 2005
Publication title -
biotechniques/biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/000112013
Subject(s) - saturated mutagenesis , mutagenesis , site directed mutagenesis , primer (cosmetics) , genetics , computational biology , biology , directed evolution , gene , directed mutagenesis , mutation , mutant , chemistry , organic chemistry
Several primer prediction programs have been developed for a variety of applications. However, none of these tools allows the prediction of a large set of primers for whole gene site-directed mutagenesis experiments using the megaprimer method. We report a novel primer prediction tool (insilico.mutagenesis), accessible at www.insilico.uni-duesseldorf.de , developed for the application to high-throughput mutagenesis used in directed evolution or structure-function dependency projects, which involve the subsequent mutagenesis of a large number of amino acid positions (e.g., in whole gene saturation or gene scanning mutagenesis experiments). Furthermore, the program is suitable for all site-directed (saturation) mutagenesis approaches, such as saturation mutagenesis of promoter sequences and other types of untranslated intergenic regions. In anticipation of downstream cloning steps, the primer design tool also includes a restriction site control feature alerting the user if unwanted restriction sites have been introduced within the mutagenesis primer. The use of our tool promises to speed up the process of site-directed mutagenesis, as it instantly allows predicting a large set of primers.