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Leishmania pp 123-140 | Cite as

Cosmid Library Construction and Functional Cloning

  • Joachim ClosEmail author
  • Dorothea Zander-Dinse
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1971)

Abstract

Cosmid libraries can represent an entire genome in a library of circular DNA molecules, allowing for the faithful amplification, cloning and isolation of large genomic DNA fragments. Moreover, using the so-called shuttle cosmid vectors, genomic DNA may be propagated in bacteria and in eukaryotic cells, which is a prerequisite for classic functional cloning and for the newly described Cos-Seq strategies.

Key words

High-quality gDNA extraction Cosmid library preparation Cosmid library evaluation Functional cloning 

Notes

Acknowledgments

We thank former coworkers Cornelia Hoyer, Katja Mellenthin, Andrea Nühs, Paloma Tejera Nevado, and Eugenia Bifeld for developing the technologies, John Kelly for the gift of pcosTL cosmid, and former and current members of the research group for discussions and suggestions.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Bernhard Nocht Institute for Tropical MedicineHamburgGermany

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