OverFlap PCR : A reliable approach for generating plasmid DNA libraries containing random sequences without a template bias
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Date
2022-08
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Abstract
Over the decades, practical biotechnology researchers have aimed to improve naturally occurring proteins and create novel ones. It is widely recognized that coupling protein sequence randomization with various effect screening methodologies is one of the most powerful techniques for quickly, efficiently, and purposefully acquiring these desired improvements. Over the years, considerable advancements have been made in this field. However, developing PCR-based or template-guided methodologies has been hampered by resultant template sequence biases. Here, we present a novel whole plasmid amplification-based approach, which we named OverFlap PCR, for randomizing virtually any region of plasmid DNA without introducing a template sequence bias.
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Publisher Copyright: © 2022 Linars et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Keywords
2.8 Environmental biotechnology, 1.6 Biological sciences, 3.4 Medical biotechnology, 1.1. Scientific article indexed in Web of Science and/or Scopus database, General
Citation
Linars, A, Silamikelis, I, Gudra, D, Roga, A & Fridmanis, D 2022, 'OverFlap PCR : A reliable approach for generating plasmid DNA libraries containing random sequences without a template bias', PloS one, vol. 17, e0262968. https://doi.org/10.1371/journal.pone.0262968