Bacterial transformation
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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
