Study of Transformation Methods for Chemically Competent E. coli Cells

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dc.contributor.advisorSarıgül, İsmail, juhendaja
dc.contributor.advisorMaiväli, Ülo
dc.contributor.authorAgbakwa, Uchechi Martina
dc.contributor.otherTartu Ülikool. Loodus- ja täppisteaduste valdkond
dc.contributor.otherTartu Ülikool. Tehnoloogiainstituut
dc.date.accessioned2024-06-17T07:10:15Z
dc.date.available2024-06-17T07:10:15Z
dc.date.issued2024
dc.description.abstractBacterial transformation is the ability of bacteria to uptake foreign DNA. High transformation efficiencies (TEs) are important in various molecular cloning processes, DNA library construction and transformation of large plasmids. Therefore, common laboratory strains are made competent by treating their membranes to enhance plasmid uptake. Transformation and storage solution (TSS) is a buffer for making common laboratory E. coli competent cells. (Chung et al., 1989). Two recent studies have reported one to two-log improved TEs (Yang, Liu, et al., 2022; Yang, Yu, et al., 2022), using slightly modified transformation buffers (TSS or TSS-HI) and methods (KCM or Heat shock). Here, we intend to reproduce these two improved methods. We prepared chemically competent E. coli cells using TSS or TSS-HI buffers. The competent cells were transformed using KCM, heat shock, or heat shock + KCM methods. Results revealed that E. coli strains washed with the modified TSS and TSS-HI buffers and transformed via different methods did not exhibit any significant improvement or difference in the transformation efficiency. However, the modified protocols produce competent cells suitable for quick transformation, eliminating the need for incubation on ice (usually 30 minutes) or recovery (about 1 hour). Our findings refute the reported modifications' effectiveness, reliability, and reproducibility in improving TEs.
dc.identifier.urihttps://hdl.handle.net/10062/99627
dc.language.isoen
dc.publisherTartu Ülikool
dc.rightsAttribution-NonCommercial-NoDerivs 3.0 Estoniaen
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/3.0/ee/
dc.subjectBacterial transformation
dc.subjectTransformation efficiency
dc.subjectCompetent cells
dc.subject.otherbakalaureusetöödet
dc.titleStudy of Transformation Methods for Chemically Competent E. coli Cells
dc.title.alternativeUuring keemiliselt kompetentsete E. coli rakkude transformatsioonimeetodite kohta
dc.typeThesis

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Science and Technology - Bachelor's theses. Kuni 2024