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Efficient preparation of shuffled DNA libraries through recombination (Gateway) cloning

Research output: Contribution to journalArticleScientificpeer-review

Details

Original languageEnglish
Pages (from-to)23-28
Number of pages6
JournalProtein Engineering Design and Selection
Volume28
Issue number1
DOIs
Publication statusPublished - 2015
Publication typeA1 Journal article-refereed

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 DNAshuffled 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.

Keywords

  • Directed evolution, DNA library, DNA shuffling, Phage display recombination cloning