Difference between revisions of "Lidstrom:Gibson Assembly"

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Back to [[Lidstrom:Protocols]]
Back to [[Lidstrom:Protocols]]
== Janet's Tips ==
'''Janet is developing a protocol [[Janet B. Matsen:Guide to Gibson Assembly |here]]'''

Revision as of 13:15, 22 March 2013

Back to Lidstrom:Protocols

Janet's Tips

Janet is developing a protocol here



  • overlap by 40 bp
  • efficiency

Oligo (primer) Design




  • Electroporation can give much higher efficiency. For this reason, the assembled product is usually electroporated.
    • rxn product is salty: can be a problem for electrocompetent cells. Rob Egbert dilutes the competent cells when transforming with electroporation. Not so much a problem when using chemically competent cells.
    • Janet hasn't had a problem transforming 1 uL of product if a cuvette is new. Older cuvettes arc more easily.


  • Colony PCR 1st
  • Sequence a few clones with primers that cover the "seams."


- Electrocompetence is ~ 1000x more effective so is tempting. -