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    This document summarizes a student's fall 2022 semester, focusing on their learning of laboratory techniques including microbiology, PCR, gel electrophoresis, and electroporation. It discusses the student's experience serially diluting the bacteria Thioclava to understand its growth rate and using electroporation to insert DNA into electrocompetent Thioclava cells. It also provides a brief history of electroporation, noting early studies in the 1970s and 1980s that demonstrated electric pulses could make cell membranes more permeable without destroying them.

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    This document summarizes a student's fall 2022 semester, focusing on their learning of laboratory techniques including microbiology, PCR, gel electrophoresis, and electroporation. It discusses the student's experience serially diluting the bacteria Thioclava to understand its growth rate and using electroporation to insert DNA into electrocompetent Thioclava cells. It also provides a brief history of electroporation, noting early studies in the 1970s and 1980s that demonstrated electric pulses could make cell membranes more permeable without destroying them.

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    43 views16 pages

    Electroporation and Other Things

    Uploaded by

    api-654986520
    This document summarizes a student's fall 2022 semester, focusing on their learning of laboratory techniques including microbiology, PCR, gel electrophoresis, and electroporation. It discusses the student's experience serially diluting the bacteria Thioclava to understand its growth rate and using electroporation to insert DNA into electrocompetent Thioclava cells. It also provides a brief history of electroporation, noting early studies in the 1970s and 1980s that demonstrated electric pulses could make cell membranes more permeable without destroying them.

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    of 16

    Electroporation Troubleshooting

    (and Other Things)

    A Recap of my Fall ‘22 Semester


    Jackson Quinn
    Who Am I?
    ▪ Graduated from Chagrin Falls High School

    ▪ Sophomore - Environmental Engineering

    ▪ Minors in Chemical Engineering and Spanish

    ▪ Work Experience:

    ▪ Ohio EPA – Engineering Intern (Summer 2022)

    ▪ Honda Motors – EHS Co-op (Spring 2023)

    Background
    Plates, plates, plates…

    Semester
    Overview
    Learning Lab Techniques (Thanks Jin!)
    ▪ Basic Microbiology

    ▪ Aseptic technique, growing/maintaining

    cultures

    ▪ PCRs & Gel Electrophoresis

    ▪ General Understanding of Genetics

    Semester
    Overview
    Serial Plate Dilutions
    ▪ Understanding Thioclava growth rate

    ▪ Connecting absorbance to cell count

    ▪ Allows for accurate co-cultures

    Semester
    Overview
    Semester
    Overview
    Electroporation of Thioclava

    Electroporation Process

    ▪ Preparing electrocompetent cells

    ▪ Add DNA to cells

    ▪ Electroporate cells

    ▪ Allow cells to recover in growth medium


    Steward, An Introduction to Electroporation – A Tool for Transfection and Competent Cell Generation

    Electroporation
    Why did using sucrose to
    make electrocompetent
    cells work?

    Electroporation
    Electroporation of Thioclava

    Electroporation
    https://microbiologysociety.org/news/society-news/professor-allan-hamilton-1936-2021.html

    History of Electroporation
    Professor Allan Hamilton (1936-2021)

    ▪ First Chair of Microbiology at the University of

    Aberdeen

    A.J.H Sale

    ▪ Very little information

    Journal Review
    History of Electroporation
    ▪ First paper suggesting changes to the

    membrane after an electrical pulse is applied

    ▪ Proved that lysis of the cells was caused by the

    electric field, not from heating

    Journal Review
    History of Electroporation

    Journal Review
    History of Electroporation
    ▪ Membrane structures can be made more

    permeable by applying an electric pulse without

    destroying the membranes

    ▪ Electric impulses can not only induce exchange

    of material across the membrane but can also

    cause fusion

    Journal Review
    History of Electroporation
    ▪ Developed some crucial knowledge of Electroporation

    ▪ Incubation time

    ▪ DNA concentration

    ▪ Time interval between pulses

    ▪ Strength of pulses

    Journal Review
    Pulse Strength
    History of Electroporation

    DNA Conc.

    Journal Review
    References & Acknowledgements

    ▪ Thanks to everyone in the lab for your

    continued help and support throughout the

    semester!

    ▪ Neumann E, Schaefer-Ridder M, Wang Y, Hofschneider PH. Gene transfer into mouse lyoma cells by electroporation in high electric fields. EMBO J.

    1982;1(7):841-5. doi: 10.1002/j.1460-2075.1982.tb01257.x. PMID: 6329708; PMCID: PMC553119.

    ▪ A.J.H. Sale, W.A. Hamilton, Effects of high electric fields on micro-organisms: III. Lysis of erythrocytes and protoplasts, Biochimica et Biophysica Acta (BBA) -

    Biomembranes, Volume 163, Issue 1, 1968, Pages 37-43, ISSN 0005-2736, https://doi.org/10.1016/0005-2736(68)90030-8.
    References

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