Figure 2. Confirmation of clones by double-digestion. The 2% agarose gel electrophoresis image shows confirmation of clones by DNA double-digestion using the restriction enzymes NcoI and BamHI. Lanes 2, 4, 6, and 8 represent the undigested plasmid of the clones R12C, R21L, R49C, and R54C mutants. Lanes 3, 5, 7, and 9 denote the double-digested products of the R12C, R21L, R49C, and R54C mutants. The release of the insert on double-digestion was confirmed by the appearance of a distinct band at approximately 512 bp.