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. Author manuscript; available in PMC: 2010 May 1.
Published in final edited form as: Methods. 2009 Feb 24;48(1):54–62. doi: 10.1016/j.ymeth.2009.02.012

Figure 4.

Figure 4

Schematic structure of the recoverable dual mutation-reporter construct p2RT. The non-B DNA forming sequences (shown as “Non-B”), or the control sequence can be inserted in either the supF gene or the lacZ gene (as shown in figure). The additional LacI binding site (shown as “I” in the figure), and an original one in Plac are used to pull down the plasmid DNA from the genomic DNA using the LacI protein coupled to magnetic beads. Plasmid DNA can be linearized by EcoRV digestion so that the vector fragment in the genome is flanked by SpeI sites on each side to facilitate isolation of the plasmid DNA from the genomic DNA, and religation for mutation screening.