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    發布時間:2019-05-21 21:53 原文鏈接: Bacterialtransformation

    Introduction

    Transformation is the process of introducing foreign DNA (e.g plasmids, BAC) into a bacterium. Bacterial cells into which foreign DNA can be transformed are called competent. Some bacteria are naturally competent (e.g B. subtilis), whereas others such as E. coli are not naturally competent. Non-competent cells can be made competent and then transformed via one of two main approaches; chemical transformation and electroporation.

    There are advantages and disadvantages to both transformation methods. In general, chemical transformation is less prone to error and faster however electroporation produces a higher transformation efficiency (fraction of transformed cells that actually uptake the foreign DNA). See Molecular Cloning for a fuller discussion of both approaches.

    Protocols

    OpenWetWare already has a number of protocols relating to bacterial transformation but more are always welcome.

    If you use a variant on one of these protocols please feel free to add a link to your protocol from one of these pages so other users can find a protocol that works for them. Additionally, if anyone uses the Innoe or Hanahan high-efficiency protocols, then please add protocols here.

    Chemical transformation

    If you plan on doing a chemical transformation, then you should see these pages -

    • Preparing chemically competent cells

    • Preparing TSS buffer

    • Transforming chemically competent cells

    • Preparing chemically competent cells (Inoue)

    • Transforming chemically competent cells (Inoue)

    • TOP10 chemically competent cells

    Chemical transformation buffer comparison

    Someone should check out the claims of Nishimura90. tk 08:58, 25 September 2007 (EDT)

    Rubidium chloride transformation protocol here


    Someone should check the claims of 1e10 chemical competence using 10% ethanol and calcium chloride protocols here.

    Electroporation

    If you plan on using electroporation, then see these pages -

    • Electrocompetent cells

    • Electroporation


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    色av