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Mesoplasma florum simplification Main Page
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Part cloning

  • J70023, J70023-F2 into pSB1AK3
  • J70024, J70025 into pSB1AC3
  • J70026 into pSB1AK3
  • J70007 into pSB1AK3

Backbone construction

  • PCR, column cleanup (100 ul volume, elute to 50 ul, digest at 100 ul)
  • digest, heat kill, add 150 ul beads, add 500 mM salt, 1 mM EDTA, volume at 276 ul)
  • bind, remove beads
  • dilute with 5x PB (1.4 ml), column bind total of 1.7 ml, 3x column loading
  • elute

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