Implementation of semi-automated cloning and prokaryotic expression screening: the impact of SPINE.

@article{Alzari2006ImplementationOS,
  title={Implementation of semi-automated cloning and prokaryotic expression screening: the impact of SPINE.},
  author={Pedro M Alzari and Helena Berglund and Nick S. Berrow and Elena V. Blagova and Didier Busso and Christian Cambillau and Val{\'e}rie Campanacci and Evangelos Christodoulou and Sylvia Eiler and Mark J. Fogg and Gerd Folkers and Arie Geerlof and Daniel A. Hart and Ahmed Haouz and Maria Dolores Herman and Sofia Macieira and Paer Nordlund and Anastassis Perrakis and Sophie Quevillon-Ch{\'e}ruel and F Tarandeau and Herman van Tilbeurgh and Tamar Unger and Mark P A Luna-Vargas and Marissa Velarde and M Willmanns and Raymond J. Owens},
  journal={Acta crystallographica. Section D, Biological crystallography},
  year={2006},
  volume={62 Pt 10},
  pages={
          1103-13
        }
}
The implementation of high-throughput (HTP) cloning and expression screening in Escherichia coli by 14 laboratories in the Structural Proteomics In Europe (SPINE) consortium is described. Cloning efficiencies of greater than 80% have been achieved for the three non-ligation-based cloning techniques used, namely Gateway, ligation-indendent cloning of PCR products (LIC-PCR) and In-Fusion, with LIC-PCR emerging as the most cost-effective. On average, two constructs have been made for each of the… CONTINUE READING

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