IGEM:IMPERIAL/2007/Projects/Cell by date/Implementation/Calibrating Absorbance: Difference between revisions
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(New page: =Calibrating Absorbance= ===Summary=== ===Materials Required=== ====Equipment==== #Spectrophotometer set to wavelength 600nm #Cuvettes ====Reagent==== ===Protocol=== #To remove...) |
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Revision as of 07:54, 2 August 2007
Calibrating Absorbance
Summary
Materials Required
Equipment
- Spectrophotometer set to wavelength 600nm
- Cuvettes
Reagent
Protocol
- To remove interference that M9 medium has with culture readings, take a reading of the broth alone ('blank'). To do this, Add 4ml of M9 broth alone to cuvette and record down the readings at 600nm.
- Next, take absorbance readings at a wavelength of 600nm for each time point, and record the relative turbidity of the broth.
- Subtract the 'blank' to measure actual OD600 of culture.
- Convert readings to CFU with calibration curve and record in representative table below.
Treatment 1: M9, 4°C Blank OD =
Time Point | Visible Observation | OD600 (abs units) | Number of Cells(CFU/ml) | |