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. Author manuscript; available in PMC: 2020 Aug 22.
Published in final edited form as: Mol Cell. 2019 Aug 22;75(4):700–710.e6. doi: 10.1016/j.molcel.2019.07.032

Figure 6. MORC-1 selectively retains circular DNA after high salt washes.

Figure 6.

(A) MORC-1 was incubated with DNA of varying topologies and precipitated with magnetic resin, then washed twice before proteinase K elution. Retained DNA is visualized by resolving samples on an 1% 1x TAE agarose gel and post-stained with ethidium bromide. (B) 400 nM MORC-1 was incubated with 50 ng open circular DNA that was made by Nt.BsmAI digestion and precipitated with magnetic resin, then washed twice with a high salt buffer and once with Restriction Enzyme (RE) buffer before digesting with restriction enzymes to alter the topology. The sample was then washed using low or high salt, then eluted using proteinase K. Retained DNA is visualized by resolving samples on an 1% 1x TAE agarose gel and post-stained with ethidium bromide. Note that PstI digestion was not complete and there were trace amounts of open circular plasmid remaining. However, MORC-1 washed with high salt only retains the open circular form and not the linearized material.

See also Figure S6.