OverFlap PCR: A reliable approach for generating plasmid DNA libraries containing random sequences without a template bias

Artis Linars, Ivars Silamikelis, Dita Gudra, Ance Roga, Davids Fridmanis (Corresponding Author)

Research output: Contribution to journalArticlepeer-review

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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.

Original languageEnglish
Article numbere0262968
Number of pages22
JournalPloS one
Volume17
DOIs
Publication statusPublished - Aug 2022
Externally publishedYes

Field of Science*

  • 2.8 Environmental biotechnology
  • 1.6 Biological sciences
  • 3.4 Medical biotechnology

Publication Type*

  • 1.1. Scientific article indexed in Web of Science and/or Scopus database

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