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