Article
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Understanding the Different Parameters That Affects Bacterial Transformation Efficiency
Version 1
: Received: 11 May 2023 / Approved: 11 May 2023 / Online: 11 May 2023 (11:49:13 CEST)
How to cite: Ng, W. Understanding the Different Parameters That Affects Bacterial Transformation Efficiency. Preprints 2023, 2023050849. https://doi.org/10.20944/preprints202305.0849.v1 Ng, W. Understanding the Different Parameters That Affects Bacterial Transformation Efficiency. Preprints 2023, 2023050849. https://doi.org/10.20944/preprints202305.0849.v1
Abstract
Bacterial transformation is the essential tool for introducing foreign DNA into bacterial cells in a host of recombinant technology applications useful for biomanufacturing and drug production. Thus far, chemically competent cells remain an important enabling technology for bacterial transformation given its facile transformation approach. However, many parameters affect transformation efficiency, and there is relatively little documentation of their cross-interacting effect in the literature. This work explores the roles played by incubation time on ice, competent cell concentration, and plasmid-to-cell ratio on transformation efficiency in Escherichia coli DH5α. Results revealed that increases in incubation time improve transformation efficiency, but 2 minutes incubation still yield appreciable efficiency. On the other hand, low concentration competent cells (106 CFU/mL) yielded significant improvement in transformation efficiency compared to high concentration competent cells (108 CFU/mL). In terms of plasmid-to-cell ratio, higher ratio increases transformation efficiency. Overall, bacterial transformation is incompletely understood particularly in the area of cross-interacting parameters. Experiments reported here revealed the possibility of short duration transformation where lower competent cell concentration and higher plasmid-to-cell ratio could improve transformation.
Keywords
Escherichia coli; bacterial transformation; plasmid-to-cell ratio; competent cell concentration; incubation time
Subject
Biology and Life Sciences, Biology and Biotechnology
Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
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