Important notes: DIMPLE breaks a gene up into sub-library fragments and generates mutagenic insert oligo pools, where each oligo contains barcodes, Type IIS restriction cutsites, and a sub-region of the gene. Be sure to review your library generation vector and gene sequences and look for pre-existing Type IIS restriction sites. Use site-directed mutagenesis to remove unwanted off-target sites. Also note that, by default, DIMPLE expects restriction enzymes with 6 bp recognition sites and 4 bp overhang lengths (as with BsaI and BsmBI).