FosmidMAX™ DNA Purification Kit

Applications

• Isolation of exceptionally pure fosmid DNA that can be used for many applications including sequencing, fingerprinting, PCR, and preparation of shotgun libraries.

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 Epicentre’s 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.

Benefits

• Simple protocols isolate up to 0.6, 15, or 25 µg of fosmid DNA from 1.5-, 40-, or 100-ml cultures of a single-copy fosmid, respectively.
• The FosmidMAX kit does not use columns or binding resins that reduce yields and shear the fosmid DNA.
• Isolate the amount of DNA that is needed, without wasting kit reagents.
• Ideal for isolating DNA from single-copy fosmid clones, induced CopyControl fosmid and cosmid clones.
• No columns or toxic organic solvents.

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.

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