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Molecular Cloning Kits

Molecular Cloning Kits: Cloning Kits

FosmidMAX™ DNA Purification Kit

The FosmidMAX™ DNA Purification Kit was developed for easy, reliable isolation of high-quality fosmid DNA

BioSearch Tech (Lucigen/Epicentre)

Description

Easy, reliable isolation of high-quality fosmid DNA

  •  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, with an easy to use protocol
  • Maximize yields because the FosmidMAX kit does not use columns or binding resins that reduce yields
  • Reduce costs by using only the reagents you need based on the amount of DNA being purified
  • Purify DNA from single-copy fosmid clones, induced CopyControl fosmid and cosmid clones
  • Reduce waste and use of hazardous reagents because no columns or toxic organic solvents are used

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 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

 




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Protocols

Protocols for: FosmidMax™ DNA Purification Kit

Due to the constant updating of the protocols by the manufacturer we have provided a direct link to Epicentre’s product page, where the latest protocol is available.

Please note this will open a new page or window on your computer.

FosmidMax™ Protocol

(catalogue number FMAX046)

Please note: all protocols off site are the responsibility of the products supplier

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References

References:

  1. Marin, M., et al. (2010) Modified 3-Oxoadipate Pathway for the Biodegradation of Methylaromatics in Pseudomonas reinekei MT1, J. Bacteriol. 192 , 1543-1552.
  2. Nishino, S. F., et al. (2010) Growth of bacteria on 3-nitropropionic acid as a sole carbon, nitrogen and energy source, Appl. Envir. Microbiol. , AEM.00267-10.
  3. Qu, Y. & Spain, J. C. (2010) Biodegradation of 5-Nitroanthranilic Acid by Bradyrhizobium sp. Strain JS329, Appl. Envir. Microbiol. 76 , 1417-1422.
  4. Camara, B., et al. (2009) Characterization of a gene cluster involved in 4-chlorocatechol degradation by Pseudomonas reinekei MT1, J. Bacteriol. , JB.00331-09.
  5. Marin, M., et al. (2009) Crystal Structure and Catalytic Mechanism of 4-Methylmuconolactone Methylisomerase, J. Biol. Chem. 284 , 32709-32716.
  6. Camara, B., et al. (2007) A Gene Cluster Involved in Degradation of Substituted Salicylates via ortho Cleavage in Pseudomonas sp. Strain MT1 Encodes Enzymes Specifically Adapted for Transformation of 4-Methylcatechol and 3-Methylmuconate, J. Bacteriol. 189 , 1664-1674.

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200

Applications & Benefits

Applications

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

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.

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200

Related products

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200