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Publication : Introduction of the negative selection marker into replacement vectors by a single ligation step.

First Author  Tzimagiorgis G Year  1996
Journal  Nucleic Acids Res Volume  24
Issue  17 Pages  3476-7
PubMed ID  8811108 Mgi Jnum  J:86581
Mgi Id  MGI:2680819 Doi  10.1093/nar/24.17.3476
Citation  Tzimagiorgis G, et al. (1996) Introduction of the negative selection marker into replacement vectors by a single ligation step. Nucleic Acids Res 24(17):3476-7
abstractText  Gene targeting is a powerful method for introducing mutations into the genome of embryonic stem cells. The most widely used approach is the positive-negative selection method in which a gene encoding a negative selection marker is cloned into the replacement vector to obtain an enrichment of properly targeted clones. Here, we present an alternative means to introduce any given negative selection marker at the ends of a replacement vector using a single ligation step, thereby avoiding laborious cloning procedures. Our results demonstrate that this fast and simple method consistently provides a high level of enrichment of appropriately targeted clones.
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