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Characterization of genome-wide ordered sequence-tagged Mycobacterium mutant libraries by Cartesian Pooling-Coordinate Sequencing

Kristof Vandewalle (UGent) , Nele Festjens (UGent) , Evelyn Plets (UGent) , Marnik Vuylsteke, Yvan Saeys (UGent) and Nico Callewaert (UGent)
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Organization
Abstract
Reverse genetics research approaches require the availability of methods to rapidly generate specific mutants. Alternatively, where these methods are lacking, the construction of pre-characterized libraries of mutants can be extremely valuable. However, this can be complex, expensive and time consuming. Here, we describe a robust, easy to implement parallel sequencing-based method (Cartesian Pooling-Coordinate Sequencing or CP-CSeq) that reports both on the identity as well as on the location of sequence-tagged biological entities in well-plate archived clone collections. We demonstrate this approach using a transposon insertion mutant library of the Mycobacterium bovis BCG vaccine strain, providing the largest resource of mutants in any strain of the M. tuberculosis complex. The method is applicable to any entity for which sequence-tagged identification is possible.
Keywords
DNA, TRANSPOSON, TUBERCULOSIS, ESSENTIAL GENES

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Please use this url to cite or link to this publication:

MLA
Vandewalle, Kristof et al. “Characterization of Genome-wide Ordered Sequence-tagged Mycobacterium Mutant Libraries by Cartesian Pooling-Coordinate Sequencing.” NATURE COMMUNICATIONS 6 (2015): n. pag. Print.
APA
Vandewalle, K., Festjens, N., Plets, E., Vuylsteke, M., Saeys, Y., & Callewaert, N. (2015). Characterization of genome-wide ordered sequence-tagged Mycobacterium mutant libraries by Cartesian Pooling-Coordinate Sequencing. NATURE COMMUNICATIONS, 6.
Chicago author-date
Vandewalle, Kristof, Nele Festjens, Evelyn Plets, Marnik Vuylsteke, Yvan Saeys, and Nico Callewaert. 2015. “Characterization of Genome-wide Ordered Sequence-tagged Mycobacterium Mutant Libraries by Cartesian Pooling-Coordinate Sequencing.” Nature Communications 6.
Chicago author-date (all authors)
Vandewalle, Kristof, Nele Festjens, Evelyn Plets, Marnik Vuylsteke, Yvan Saeys, and Nico Callewaert. 2015. “Characterization of Genome-wide Ordered Sequence-tagged Mycobacterium Mutant Libraries by Cartesian Pooling-Coordinate Sequencing.” Nature Communications 6.
Vancouver
1.
Vandewalle K, Festjens N, Plets E, Vuylsteke M, Saeys Y, Callewaert N. Characterization of genome-wide ordered sequence-tagged Mycobacterium mutant libraries by Cartesian Pooling-Coordinate Sequencing. NATURE COMMUNICATIONS. 2015;6.
IEEE
[1]
K. Vandewalle, N. Festjens, E. Plets, M. Vuylsteke, Y. Saeys, and N. Callewaert, “Characterization of genome-wide ordered sequence-tagged Mycobacterium mutant libraries by Cartesian Pooling-Coordinate Sequencing,” NATURE COMMUNICATIONS, vol. 6, 2015.
@article{6887749,
  abstract     = {Reverse genetics research approaches require the availability of methods to rapidly generate specific mutants. Alternatively, where these methods are lacking, the construction of pre-characterized libraries of mutants can be extremely valuable. However, this can be complex, expensive and time consuming. Here, we describe a robust, easy to implement parallel sequencing-based method (Cartesian Pooling-Coordinate Sequencing or CP-CSeq) that reports both on the identity as well as on the location of sequence-tagged biological entities in well-plate archived clone collections. We demonstrate this approach using a transposon insertion mutant library of the Mycobacterium bovis BCG vaccine strain, providing the largest resource of mutants in any strain of the M. tuberculosis complex. The method is applicable to any entity for which sequence-tagged identification is possible.},
  articleno    = {7106},
  author       = {Vandewalle, Kristof and Festjens, Nele and Plets, Evelyn and Vuylsteke, Marnik and Saeys, Yvan and Callewaert, Nico},
  issn         = {2041-1723},
  journal      = {NATURE COMMUNICATIONS},
  keywords     = {DNA,TRANSPOSON,TUBERCULOSIS,ESSENTIAL GENES},
  language     = {eng},
  pages        = {7},
  title        = {Characterization of genome-wide ordered sequence-tagged Mycobacterium mutant libraries by Cartesian Pooling-Coordinate Sequencing},
  url          = {http://dx.doi.org/10.1038/ncomms8106},
  volume       = {6},
  year         = {2015},
}

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