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Isolation of Cosmid and BAC DNA from E. coli

  • Daniel Sinnett
  • Alexandre Montpetit
Part of the Methods in Molecular Biology™ book series (MIMB, volume 235)

Abstract

Cosmid and bacterial artificial chromosome (BAC) systems have been developed for the cloning of large DNA inserts averaging 40 kb and 130 kb (range: 90–300 kb), respectively. The resulting clones are more stable than yeast artificial chromosomes (YACs) and rarely chimeric, which makes them excellent tools for the generation of contiguous physical maps. The use of such large-insert clones considerably increases the rate of complex genome mapping and sequencing. Numerous standard protocols have been developed to isolate supercoiled plasmids, but of these, the alkaline lysis protocol remains the most suitable approach to isolate large-insert clones such as cosmids and BACs.

Keywords

Sodium Dodecyl Sulfate Bacterial Artificial Chromosome Bacterial Artificial Chromosome Clone Isoamyl Alcohol Yeast Artificial Chromosome 
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.

References

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    Birnboim, H. C. and Doly, J. (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 7, 1513–1523.PubMedCrossRefGoogle Scholar
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    Thomas, N. R., Koshy, S., Simsek, M. et al. (1988) A precaution when preparing very large plasmids by alkaline lysis procedure. Biotechnol. Appl. Biochem. 10, 402–407.PubMedGoogle Scholar
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    Sinnett, D., Richer, C., and Baccichet, A. (1998) Isolation of stable bacterial artificial chromosome DNA using a modified alkaline lysis method. Biotechniques 24, 752–754.PubMedGoogle Scholar

Copyright information

© Humana Press Inc. 2003

Authors and Affiliations

  • Daniel Sinnett
    • 1
  • Alexandre Montpetit
    • 1
  1. 1.Service dďHùmatologie-Oncologie, Centre de Cancùrologie Charles Bruneau, Centre de Recherche, Hôpital Ste-Justine, Dùpartement de pùdiatrieUniversitù de MontrùalMontrùalCanada

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