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Topo IV (from Streptococcus pneumoniae) is prepared by overexpressing the subunits in E. coli and purifying them by methods developed in-house.
The subunits are purified to >95% purity as judged by SDS-PAGE. The topo IV is supplied as a heterotetramer complex in Dilution buffer.
It is recommended that the enzyme is aliquoted to avoid repeated freeze-thaw cycles. Store at -80ºC.
All enzyme is supplied with 5X concentrated Assay Buffer and Dilution buffers which are also available separately.
See technical documents below for more detailed information and lot specific activities.
These contain S. pneumoniae topo IV and the supercoiled DNA substrate in addition to the Assay and Dilution buffers for relaxation reactions. 1 U of topo IV will relax 0.5 µg supercoiled pBR322 DNA in 30 minutes at 37°C.
These contain S. pneumoniae topo IV and the catenated kDNA substrate in addition to the Assay and Dilution buffers for decatenation reactions. 1 U of topo IV will decatenate 200 ng of kDNA when incubated in 1X Assay buffer in a total reaction volume of 30 µl at 37°C for 30 minutes.