Bacterial transformation

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==Protocols==
==Protocols==
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OpenWetWare already has a number of protocols relating to bacterial transformation '''but more are always welcome'''.  If you plan on doing a chemical transformation, then you should see these pages -  
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OpenWetWare already has a number of protocols relating to bacterial transformation '''but more are always welcome'''.
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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 themAdditionally, if anyone uses the Innoe or Hanahan high-efficiency protocols, then please add protocols here.
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===Chemical transformation===
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If you plan on doing a chemical transformation, then you should see these pages -  
*[[Preparing chemically competent cells]]
*[[Preparing chemically competent cells]]
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*[[TOP10 chemically competent cells]]
*[[TOP10 chemically competent cells]]
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[[Chemical transformation buffer comparison]]
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===Electroporation===
If you plan on using electroporation, then see these pages -  
If you plan on using electroporation, then see these pages -  
*[[Electrocompetent cells]]
*[[Electrocompetent cells]]
*[[Electroporation]]
*[[Electroporation]]
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-
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.
 
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[[Chemical transformation buffer comparison]]
 
==References==
==References==
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{{:Molecular Cloning}}
 
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<biblio>
<biblio>
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#MolecularCloning isbn=0-87969-577-3
# Hanahan91 pmid=1943786
# Hanahan91 pmid=1943786
# Hanahan89 US Patent 4,851,348 [[Media:pat4851348.pdf]]
# Hanahan89 US Patent 4,851,348 [[Media:pat4851348.pdf]]

Revision as of 13:54, 13 May 2006

Back to Protocols

Contents

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 -

Chemical transformation buffer comparison

Electroporation

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

References

  1. isbn:0-87969-577-3. [MolecularCloning]
  2. Hanahan D, Jessee J, and Bloom FR. . pmid:1943786. PubMed HubMed [Hanahan91]
  3. US Patent 4,851,348 Media:pat4851348.pdf [Hanahan89]
  4. US Patent 4,981,797 Media:pat4981797.pdf [Jessee90]
  5. US Patent 6,247,369 Media:pat6274369.pdf [Donahue01]
  6. US Patent 6,706,525 Media:pat6706525.pdf [Greenr04]
  7. US Patent 6,709,854 Media:pat6709854.pdf [Donahue04]
  8. US Patent 6,855,494 Media:pat6855494.pdf [Bloom05]
  9. US Patent 6,960,464 Media:pat6960464.pdf [Jessee05]
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