Improved PCR-walking for large-scale isolation of plant T-DNA borders.

  title={Improved PCR-walking for large-scale isolation of plant T-DNA borders.},
  author={S. Balzergue and B. Dubreucq and S. Chauvin and I. Le-Clainche and F. le Boulaire and R. De Rose and F. Samson and V. Biaudet and A. Lecharny and C. Cruaud and J. Weissenbach and M. Caboche and L. Lepiniec},
  volume={30 3},
          496-8, 502, 504
only one sequencing reaction. Instead of using ethanol precipitation after the first PCR, one can use filtration methods in a 96-well format with QIAquick 96 (Qiagen GmbH, Hilden, Germany) or NucleoSpin Multi-96 (Macherey-Nagel GmbH, Düren, Germany). These methods remove the PCR primers more efficiently and resulted in a remarkable reduction of amplification peaks in the intron sequences. However, the quality of exon sequences was the same compared to the purification by ethanol precipitation… Expand
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