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Does anyone know the optimum length of overlap your primers should have to your vector for gibson assembly. Most manufacturers say to do at least 20 bp on both side but don't give a maximum. Sometimes I add 30-35 bp on both I don't know if that hinders efficiency.

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  • $\begingroup$ Increasing overlap should not hinder. However, you'll waste money in synthesizing more oligoes. $\endgroup$
    – WYSIWYG
    May 2, 2016 at 5:14
  • $\begingroup$ There has to be some diminishing return. $\endgroup$ May 2, 2016 at 7:17

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