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Lox-Stop-Lox TOPO Notes

Addgene diagnostic digest

This gel depicts the NEB 1kb ladder and a digestion of the plasmid with the following enzyme: XbaI

We expected the following band sizes for this digest: 1.2kb and 8.0kb. The plasmid is only linearized. However, upon examining the sequence, there appears to be an overlapping Dam methylation site at the site of the first XbaI site. It appears as though Dam methylation blocks cleavage at this XbaI site.