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. Author manuscript; available in PMC: 2011 Apr 6.
Published in final edited form as: Biochemistry. 2010 Apr 6;49(13):2786–2795. doi: 10.1021/bi902204v

Figure 2.

Figure 2

Purification of the vTopo I-DNA covalent complex. (A) Purification scheme. The enzyme is added to the biotinylated DNA duplex (dsDNA-bio) and following formation of the non-covalent complex (NCC-bio), the covalent complex is formed (CC) and the biotinylated single strand (ssDNA-bio) dissociates from the covalent complex. The reaction mixture is incubated with streptavidin beads and filtered, resulting in separation of the biotinylated products and the purified covalent complex. FAM label used in subsequent experiments depicted with an open circle. (B) SDS-PAGE gel of purification steps. Lane 1: Biotinylated DNA duplex (dsDNA) alone. Lane 2, Reaction mixture prior to strepdavidin-agarose purification. Lane 3: Purified covalent complex.