Oligoreef - Generation of Primers for Complex Polymerase Chain Reactions

Oligoreef is a new tool to generate composite primers for complex polymerase chain reactions (PCR) in an easy and time-saving way. Various PCR techniques using composite primers can be employed to perform quick in vitro mutagenesis. For instance overlap extension PCR intending to insert particular restriction sites, also the deletion or substitution of specific nucleotide sequences are conceivable.

Oligoreef allows obtaining suitable composite primers to run PCRs for various purposes considering recommended general primer properties and adequate melting temperatures. Automatic determination of the best fitting complementary sequence, targeting matching purposes of respective primers for both directions, is processed. An application example is presented - using different fluorescent proteins, regulated by different inducible promoters, generated by overlap-extension PCR, cloned into the standard plasmid and expressed in E. coli.

Oligoreef is available online at http://hgen-oligoreef.hgen.slu.se:8080