Imperial College London/Notebook/17 September 2009

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Contents

Testing

OD calibration 17th Sep

Protocol considerations

  • Cells were grown overnight in 10ml of M9 media
  • The cells were allowed to grow until a high OD (very cloudy) and retrieved in the early afternoon
  • The OD was measured using the multiplate reader.
  • As the OD was only around 1.5, the cells were centrifuged and resuspended in less media to form an OD of 3
  • OD of around 0.5 to 3.0 was made up, and subsequently diluted up to 10-6 to 10-8
  • Plated on strep plates and dried until the end of the day
  • Placed into 37 degrees incubator (non-shaking) overnight

Results

The plates had too many colonies to count.

Conclusions

  • The plates were all too confluent, even at 10-8 dilution.
  • Unable to count colonies and draw any colonies
  • Need to repeat experiment


Suggestions/Improvements

  • Was not done on ice, so cell growth still occurred during dilutions
  • Too many colonies to count when plated 100uL
    • Use ice
    • Increase dilutions


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