Construction and Screening of Cosmid Libraries

  • Jens Hanke
  • Jörg D. Hoheisel
Part of the Springer Protocols Handbooks book series (SPH)


Cosmid libraries still are an important tool for cloning large genomic regions. Although other Escherichia coli-based vector systems like bacteriophage PI, bacterial artificial chromosomes (BACs), and PI-based artificial chromosomes (PACs) allow the cloning of longer inserts, they are more difficult to handle (1). Therefore, even if they are frequently used for the initial generation of libraries from large genomes, cosmid vectors are often still required to generate libraries with smaller, more manageable pieces or libraries from limited DNA amounts, such as bands isolated from pulsed-field gels (2).


Bacterial Artificial Chromosome Phage Particle Cosmid Library Ligation Buffer Cosmid Vector 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


  1. 1.
    Monaco, A. P. and Larin, Z. (1994) YACs, BACs, PACs and MACs: artificial chromosomes as research tools. Trends Biotechnol. 12(7), 280–286.PubMedCrossRefGoogle Scholar
  2. 2.
    Scholler, P., Schwarz, S., and Hoheisel, J. D. (1995) High-resolution cosmid mapping of the left arm of Saccharomyces cerevisiae chromosome XII; a first step towards an ordered sequencing approach. Yeast 11, 659–666.PubMedCrossRefGoogle Scholar
  3. 3.
    Sambrook, J., Fritsch, E. F., and Maniatis, T. (eds.) (1989) Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.Google Scholar
  4. 4.
    Bates, P. F. and Swift, R. A. (1983) Double cos site vectors: Simplified cosmid cloning. Gene 26, 137–146.PubMedCrossRefGoogle Scholar
  5. 5.
    Poustka, A., Rackwitz, H. R., Frischauf, A. M., Hohn, B., and Lehrach, H. (1984) Selective isolation of cosmid clones by homologous recombination in Escherichia coli. Proc. Natl Acad. Set USA 81, 4129–4133.CrossRefGoogle Scholar
  6. 6.
    Kioussis, D., Wilson, F., Daniels, C., Leveton, C., Taverne, J., and Playfair J. H. L (1987) Expression and rescuing of a cloned human tumor necrosis factor gene using an EBV-based shuttle cosmid vector. EMBO J. 6, 355–361.PubMedGoogle Scholar
  7. 7.
    de Jong, P. J., Chen, C, and Games, J. (1989) Application of PCR for the construction of vectors and the isolation of probes, in Polymerase Chain Reaction (Erlich, H. A., Gibbs, R. A., and Kazazian, H. H., eds.), Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, pp. 205–210.Google Scholar
  8. 8.
    Clarke, L. and Carbon, J. (1976) A colony bank containing synthetic ColEl hybrid plasmids representative of the entire E. coli genome. Cell 9, 91–99.PubMedCrossRefGoogle Scholar
  9. 9.
    Maier, E., Hoheisel, J. D., McCarthy, L., Mott, R., Grigoriev, A. V., Monaco, A. P., Larin, Z., and Lehrach, H. (1992) Yeast artificial chromosome clones completely spanning the genome of Schizosaccharomyces pombe. Nature Genet. 1, 273–277.PubMedCrossRefGoogle Scholar
  10. 10.
    Hoheisel, J. D. and Lehrach, H. (1993) Use of reference libraries and hybridisation fingerprinting for relational genome analysis. FEBS Lett. 325, 118–122.PubMedCrossRefGoogle Scholar
  11. 11.
    Hoheisel, J. D., Maier, E., Mott, R., and Lehrach, H. (1995) Integrated genome mapping by hybridisation techniques, in Analysis of Non-Mammalian Genomes–A Practical Guide (Birren, B. and Lai, E., eds.), Academic Press, London, pp. 319–346.Google Scholar
  12. 12.
    Larin, Z., Monaco, A. P., Meier-Ewert, S., and Lehrach, H. (1993) Construction and characterisation of yeast artificial chromosome libraries from the mouse genome. Methods Enzymol 255, 623–637.CrossRefGoogle Scholar
  13. 13.
    Lehrach, H., Drmanac, R., Hoheisel, J. D., Larin, Z., Lennon, G., Monaco, A. P., Nizetic, D., Zehetner, G., and Poustka, A. (1990) Hybridisation fingerprinting in genome mapping and sequencing, in Genome Analysis: Genetic and Physical Mapping (Davies, K. E. and Tilghman, S., eds.), Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, pp. 39–81.Google Scholar
  14. 14.
    O’Farrell, P. H., Kutter, E., and Nakanishi, M. (1980) A restriction map of the bacterioph-age-T4 genome. Mol Gen. Genet. 179, 421–435.PubMedCrossRefGoogle Scholar
  15. 15.
    Morgan, A. R., Lee, J. S., Pulleyblank, D. E., Murray, N. L., and Evans, D. H. (1979) Review: ethidium fluorescence assays. Part 1. Physicochemical studies. Nucleic Acids Res. 7(3), 547–569.PubMedCrossRefGoogle Scholar
  16. 16.
    Hoheisel, J. D., Nizetic, D., and Lehrach, H. (1989) Control of partial digestion combining the enzymes Dam methylase and Mbol. Nucleic Acids Res. 17, 4571–4582.CrossRefGoogle Scholar
  17. 17.
    Feinberg, A. P. and Vogelstein, B. (1983) A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal. Biochem. 132, 6–13.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 2000

Authors and Affiliations

  • Jens Hanke
    • 1
  • Jörg D. Hoheisel
    • 1
  1. 1.Functional Genome AnalysisDeutsches KrebsforschungszentrumHeidelbergGermany

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