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Step 2: Insertion of Fragments into a Plasmid

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All the small random fragments generated by shearing are inserted, or "ligated," into a plasmid (a loop of nonessential bacterial DNA) called pUC18. The pUC18 vector allows for propagation and replication in Escherichia coli bacteria (E. coli). Each plasmid has a gene for antibiotic resistance (green in the diagram below) as well as a LacZ gene (blue) that produces blue pigment. These two genes will be important later. The fragments, shown here in various colors, are inserted in the middle of the LacZ gene.

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