AI Article Synopsis
- Developed a more efficient method for subcloning DNA-shuffled libraries using recombination cloning (Gateway) without extra purification steps.
- The new procedure simplifies traditional DNA shuffling protocols and maintains high library quality.
- This method reduces the time needed to construct diverse DNA libraries, making it generally applicable for gene library creation.
Article Abstract
Efficient and robust subcloning is essential for the construction of high-diversity DNA libraries in the field of directed evolution. We have developed a more efficient method for the subcloning of DNA-shuffled libraries by employing recombination cloning (Gateway). The Gateway cloning procedure was performed directly after the gene reassembly reaction, without additional purification and amplification steps, thus simplifying the conventional DNA shuffling protocols. Recombination-based cloning, directly from the heterologous reassembly reaction, conserved the high quality of the library and reduced the time required for the library construction. The described method is generally compatible for the construction of DNA-shuffled gene libraries.
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| http://dx.doi.org/10.1093/protein/gzu050 | DOI Listing |
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