Janet B. Matsen:Guide to Gibson Assembly

From OpenWetWare
Revision as of 05:35, 9 October 2012 by Janet B. Matsen (talk | contribs)
Jump to: navigation, search

Back to Janet


  • 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.


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