Team:Bielefeld-CeBiTec/Notebook/Journal/Isobutanol/Jul

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July

  • alsS, ilvC, ilvD, kivD and backbone of pSB1C3
    • We tried to redo the amplification of last week.
      • Optimization of PCR conditions for coding sequence amplification
        • Combinations of primers and templates as described before
        • Annealing temperature gradients from 50°C to 58°C were tried
        • Product amount was increased by lower annealing temperatures, but still not good enough for Gibson
  • alsS, ilvC, ilvD, kivD and backbone of pSB1C3
    • We tried to redo the amplification of last week.
      • Optimization of PCR conditions for coding sequence amplification
        • Combinations of primers and templates as described before
        • Annealing temperature gradients from 50°C to 58°C were tried
        • Product amount was increased by lower annealing temperatures
          • For the annealing temperature we identified 54°C as optimal.
          • For the elongation time 90 seconds worked better than 60 seconds.