Plasmids 101: How to Verify Your Plasmid Using a Restriction Digest Analysis
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Blog Post
.... Diagnostic restriction digests are comprised of 2 separate steps: 1) incubating your DNA with restriction...which cleave the DNA molecules at specific sites and 2) running the reaction on an agarose gel to determine...including a 1.2 kb insert. The plasmid was digested with 2 unique enzymes (HindIII and BamHI) and run on an agarose...to serve as a reference band. The uncut DNA (lane 2) shows 3 possible plasmid conformations, with relaxed...Watch out for methylation issues. Enzymes like XbaI and ClaI are sensitive to methylation and their activity...will need to purify your DNA from a dcm or dam methylation-deficient bacterial strain such as JM110 or INV110...