IGEM:Cambridge/2008/Notebook/Voltage/2008/08/07

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Flame Photometry

  • Various concentrations of KCl were made up in LB to calibrate for LB instead of water.
  • Took OD600 readings of mutants grown in KCl concentrations in shaking incubator overnight
  • They were then spun down, resuspended in SDW and spun again to wash, then resuspended.
  • The resuspended cells were put through the flame photometer to discern internal cell K+ concentration.
  • The supernatant K+ concentration was also measured.

Kdp

  • Ran ligation mixtures on agarose gel and extracted 8kb (supercoiled plasmid) bands.
  • Transformed cells with cleaned up plasmid.

Mutants