Abstract
Gene synthesis of leucine zipper genes based on blunt end ligation of ds DNA modules was performed in several ligation steps on magnetic beads. A 3'-phosphate, which was removed by T4 polynucleotide kinase after each ligation step, was used to guarantee orientation and single addition of newly added modules. Not depending on sequence complementarity the method appears well suited for the generation of combinatorial libraries of heterologous molecules.
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Dietrich, R., Wirsching, F., Opitz, T. et al. Gene assembly based on blunt-ended double-stranded DNA-modules. Biotechnology Techniques 12, 49–54 (1998). https://doi.org/10.1023/A:1008855526226
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DOI: https://doi.org/10.1023/A:1008855526226