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.