The FosmidMAX™ DNA Purification Kit was developed for easy, reliable isolation of high-quality fosmid DNA (Fig. 1). The scalable protocol is based on a modified alkaline-lysis procedure that typically yields 0.6-25 µg of fosmid DNA from 1.5 to 100 ml of a single-copy fosmid culture. Selective precipitation steps and the incorporation of the Epicentre RiboShredder™ RNase Blend effectively remove contaminants that degrade DNA and interfere with downstream applications.
The FosmidMAX kit is ideal for the purification of induced fosmids produced by the CopyControl™ Fosmid Library Production Kit. The CopyControl cloning technology enables the user to grow the fosmid clones at single copy to ensure insert stability and cloning of toxic gene products and then induce the clones to high-copy number for high yields of DNA. Using the FosmidMAX Kit, up to 4 µg of fosmid DNA can be isolated from 1.5 ml of an induced clone.
|Figure 1. Quality of DNA prepared with the FosmidMAX™ Kit. Fosmid DNA isolated from six different fosmid clones (lanes 1 to 6) was digested with Hind III and analyzed by agarose gel electrophoresis. Lane M, Kilobase ladder.||Figure 2. Fosmid clones made with the CopyControl™ Fosmid Library Production Kit can be induced from single-copy up to 50 copies per cell to greatly increase DNA yield. DNA from an equal number of cells of uninduced (-) and induced (+) CopyControl fosmid clones was isolated using the FosmidMAX™ Kit. Equivalent volumes of fosmid DNA were digested with Hind III and analyzed by agarose gel electrophoresis. Lane M, kb ladder.|