CorrectASE™ Enzyme

The CorrectASE enzyme has the ability to remove mismatches caused by oligonucleotide synthesis errors and is typically used in a do-it-yourself gene synthesis workflow to aid in decreasing mutations in your synthetic genes/fragements.

The protocol states that oligonucleotide stocks should be prepared at a final concentration of 100 µM in 1X TE buffer. The next line indicates the addition of 5 µL of each 10 µM primer together. According to R&D, the manual was written this way because our R&D typically brings up the lyophilized oligos to a 100 µM stock concentration (due to the volume of the tube). You do, however, want to use a 0.15 µM pool. Therefore, you can either dilute the stock to 10 µM or dilute the primer pool 1:1

Overdigestion with CorrectASE enzyme can lead to degradation of the DNA template.

Ensure that the reaction does not go longer than 60 mins. Also, ensure that the reaction is kept on ice until the PCR step or else the reaction will be prone to overdigestion by the CorrectASE enzyme.

Please send an email to geneartsupport@lifetech.com explaining what is wrong. We will also need the project ID and construct ID, which we will forward to our QC department for further investigation.

We have a variety of strains that are used in production, such as Top10 or DH5α. Routinely, we grow them in dam+ strains. Therefore, you may be seeing inhibition because Xba1 is sensitive to dam methylation