Target sequence confirmation of M.BatI by digestion of M.BatI methylated dsDNA.

(A) Structure of 60 bp dsDNA. The site with overlapping sequences was positioned at one-third from the right end. (B) Design of overlapping sites in each dsDNA and the expected outcomes by methylation and digestion reactions. Each site included a target site of another restriction enzyme (squares). Expected positions of methylation were depicted with “Me” in a circle. (C) Digestion tests of dsDNA. Each dsDNA was first treated with M.BatI and digested with one of the four restriction enzymes.