Invitrogen™ CorrectASE™ Enzyme

Description

• CorrectASE™ enzyme removes mismatches caused by oligonuceotide synthesis errors, leading to a 3–10 fold reduction in mutations in your synthetic genes or fragments
• By introducing CorrectASE™ enzyme into your do-it-yourself gene syntheis workflow, you can: Reduce the number of mutations in your synthetic gene or fragment Reduce your labor time by screening only 2–4 clones instead of 10-16 clones per synthetic construct Reduce your costs by sequencing only 2–4 clones instead of 10–16 synthetic genes Prevent Unwanted Mutations Commercially available synthetic oligonuceotides have a high error rate during synthesis, ranging from one per 300–1000 bases, depending on the source
• These errors cause frameshift (deletion and insertion) and mismatch mutations during gene synthesis
• Incubation with CorrectASE™ enzyme removes both type of mutations
• The incubation step with CorrectASE™ enzyme is introduced after the initial PCR assembly of oligonucleotides
• The PCR product is denatured and reannealed so that any mutations will be unmatched
• CorrectASE™ enzyme binds to the resulting mismatches and nicks both DNA strands 3' of the error
• The 3' to 5' exonuclease activity of the enzyme removes the errors
• A final PCR with a proofreading polymerase then assembles the corrected fragments, thus increasing the likelihood of isolating clones with the correct sequence
• Depending upon the incoming oligonuceotide quality, only 2–4 clones need to be screened, compared to 10–16 clones in a workflow that does not include the correction step
• Including CorrectASE™ enzyme in your gene synthesis workflow decreases labor time and sequencing costs.