TY - JOUR
T1 - OverFlap PCR
T2 - A reliable approach for generating plasmid DNA libraries containing random sequences without a template bias
AU - Linars, Artis
AU - Silamikelis, Ivars
AU - Gudra, Dita
AU - Roga, Ance
AU - Fridmanis, Davids
N1 - 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.
PY - 2022/8
Y1 - 2022/8
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85135547389&partnerID=8YFLogxK
U2 - 10.1371/journal.pone.0262968
DO - 10.1371/journal.pone.0262968
M3 - Article
C2 - 35939421
AN - SCOPUS:85135547389
SN - 1932-6203
VL - 17
JO - PloS one
JF - PloS one
M1 - e0262968
ER -