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OverFlap PCR – a reliable approach for generation of plasmid DNA libraries containing random sequences without template bias

Artis Linars, Ivars Silamikelis, Dita Gudra, Ance Roga, View ORCID ProfileDavids Fridmanis
doi: https://doi.org/10.1101/2022.01.11.475799
Artis Linars
Latvian Biomedical Research and Study Centre, Riga, Latvia, LV-1067, Latvia
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Ivars Silamikelis
Latvian Biomedical Research and Study Centre, Riga, Latvia, LV-1067, Latvia
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Dita Gudra
Latvian Biomedical Research and Study Centre, Riga, Latvia, LV-1067, Latvia
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Ance Roga
Latvian Biomedical Research and Study Centre, Riga, Latvia, LV-1067, Latvia
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Davids Fridmanis
Latvian Biomedical Research and Study Centre, Riga, Latvia, LV-1067, Latvia
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  • ORCID record for Davids Fridmanis
  • For correspondence: davids@biomed.lu.lv
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ABSTRACT

Over the decades the improvement of naturally occurring proteins and creation of novel ones has been the primary goal for many practical biotechnology researchers and it is widely recognized that randomization of protein sequences coupled to various effect screening methodologies is one of the most powerful techniques for fast, efficient and purposeful approach for acquisition of desired improvements. Over the years considerable advancements have been made in this field, however development of PCR based or template guided methodologies has been hampered by the resulting template sequence bias. In this article we present novel whole plasmid amplification based approach, which we named OverFlap PCR, for randomization of virtually any region of the plasmid DNA, without introduction of mentioned bias.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
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Posted January 11, 2022.
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OverFlap PCR – a reliable approach for generation of plasmid DNA libraries containing random sequences without template bias
Artis Linars, Ivars Silamikelis, Dita Gudra, Ance Roga, Davids Fridmanis
bioRxiv 2022.01.11.475799; doi: https://doi.org/10.1101/2022.01.11.475799
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OverFlap PCR – a reliable approach for generation of plasmid DNA libraries containing random sequences without template bias
Artis Linars, Ivars Silamikelis, Dita Gudra, Ance Roga, Davids Fridmanis
bioRxiv 2022.01.11.475799; doi: https://doi.org/10.1101/2022.01.11.475799

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