Janet B. Matsen:Guide to Gibson Assembly

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(New page: * you can chose where the seam is if you use longer oligos * Dpn1 * RFP for backbone: don't screen red colonies!)
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Back to [[User:Janet B. Matsen|Janet]]
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== Intro ==
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* What is it?
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* How does it differ from other cloning?
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* When should I use it?
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* Steps (concise)
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** Design oligos to yield 40 - 100 bp overlapping linear DNA segments
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** Purify (usually gel) the PCR products (or digest)
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** Use Gibson Assembly Mix
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** Transform
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*** Electroporation is usually used to provide higher yield.
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 +
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== Examples ==
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* you can chose where the seam is if you use longer oligos
* you can chose where the seam is if you use longer oligos
* Dpn1
* Dpn1
* RFP for backbone: don't screen red colonies!
* RFP for backbone: don't screen red colonies!

Revision as of 08:35, 9 October 2012

Back to Janet

Intro

  • What is it?
  • How does it differ from other cloning?
  • When should I use it?
  • Steps (concise)
    • Design oligos to yield 40 - 100 bp overlapping linear DNA segments
    • Purify (usually gel) the PCR products (or digest)
    • Use Gibson Assembly Mix
    • Transform
      • Electroporation is usually used to provide higher yield.


Examples

  • you can chose where the seam is if you use longer oligos
  • Dpn1
  • RFP for backbone: don't screen red colonies!
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