Janet B. Matsen:Guide to Gibson Assembly

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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!