BACMAXTM or FosmidMAXTM
Fosmid DNA Purification Kit
FosmidMAX™ DNA Purification Kit
Applications
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.
- Isolation of exceptionally pure fosmid DNA that can be used for many applications including sequencing, fingerprinting, PCR, and preparation of shotgun libraries.
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.
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Figure 1. Quality of DNA prepared with the FosmidMAX™ Kit. Fosmid DNA isolated from six different fosmid clones (lanes 1 to 6) was digested withHind 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 withHind III and analyzed by agarose gel electrophoresis. Lane M, kb ladder. |
BACMAX™ DNA Purification Kit
Applications
The BACMAX™ DNA Purification Kit was developed for easy, reliable isolation of high-quality BAC DNA. The scalable protocol is based on a modified alkaline-lysis procedure that typically yields 0.6-25 µg of BAC DNA from 1.5-100 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.
- Isolation of exceptionally pure BAC DNA that can be used for many applications including sequencing, fingerprinting, PCR, and preparation of shotgun libraries.
The BACMAX Kit is ideal for the purification of induced BACs produced with CopyControl™ BAC Vectors. The CopyControl cloning technology enables the user to grow the BAC 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 BACMAX Kit, up to 4 µg of BAC DNA can be isolated from 1.5 ml of an induced clone.
Benefits
Panel A Panel B
- Simple protocols isolate up to 0.6, 15, or 25 µg of BAC DNA from 1.5-, 40- or 100-ml cultures of a single-copy BAC, respectively.
- The kit does not use columns or binding resins that lower yields and shear the DNA.
- Isolate the amount of DNA that is needed, without wasting kit reagents.
- Ideal for isolating DNA from single-copy BAC clones or induced CopyControl™ BAC clones.
- No toxic organic solvents.
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Figure 1. Obtain higher yields of BAC DNA with the BACMAX™ DNA Purification Kit. DNA was isolated from 1.5 or 3.0-ml cultures of a 153-kb single-copy BAC clone and quantitated by fluorometry with Hoechst 33258 dye. Culture volumes were those recommended by the Suppliers. |
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Figure 2. Panel A. The BACMAX™ DNA Purification Kit consistently yields high-quality BAC DNA that can be used directly for downstream applications like fingerprinting. BAC DNA isolated from 10 different clones (Lanes 1 to 10) was digested withHind III and analyzed by agarose gel electrophoresis. Lane M, kb ladder. Panel B. BAC DNA isolated with the BACMAX™ Kit shows no degradation after being stored for 1 year at –20°C. DNA isolated from a 153-kb and a 165-kb BAC clone was immediately digested with Hind III (Lane 1, 153 kb and Lane 2, 165 kb) or stored for 1 year at -20°C and then digested withHind III (Lane 3, 153 kb and Lane 4, 165 kb). Samples were analyzed by agarose gel electrophoresis. Lane M, kb ladder. |