User:Tk/Notebook/MF-xfm/2008/04/22

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Construction plasmid PCR

  • 100 ul final
  • 50 ul Phusion mastermix
  • 1.5 ul suffix-FB
  • 1.5 ul prefix-RB
  • 0.5 ul of plasmid (pSB1AC3, 1AK3, 1AT3)
  • 46.5 ul water
  • Cycle 98/30s 32x(98/20s 55/20s 72/2min) 72/10min

Transformation results

  • Picked 40 colonies off of 4/20 transformations
  • +10 worked best, 0 worked well, -5 few, -10 none, +15 few
  • replated 0,-5 saved cultures

Construction plasmid distribution

  • Each team gets 300 ul of each plasmid
  • Concentration is 10 ng/ul
  • Each team needs 3 ug of plasmid
  • With 150 teams, we need 450 ug of each plasmid


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