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. 2001 Jun 15;29(12):2492–2501. doi: 10.1093/nar/29.12.2492

Figure 4.

Figure 4

Gel retardation assay with MlyI, PleI and their variants. Conditions are described in the Figure 3 legend. Lane 1 is the size standard of λ DNA digested by HindIII, φ174 DNA digested by HaeIII. 0.25 pmol of the BssSI–BsrFI digested pNB1 was incubated with 1 µl of crude cell extracts containing ∼0.3 pmol of wild-type MlyI (lane 2) and its variants E380A (lane 3), D403A (lane 4), D418A (lane 5). Same DNA substrate (0.25 pmol) was also incubated with 1 µl of crude cell extracts containing ∼0.45 pmol of wild-type PleI and its variants D399A, D414A, D427A, shown in lanes 7, 8, 9 and 10, respectively. Lanes 6 and 11 are the controls of plasmid DNA only.