IGEM:IMPERIAL/2007/Projects/Cell by date/Implementation/Calibrating Absorbance: Difference between revisions
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#Convert readings to CFU with calibration curve and record in representative table below. | #Convert readings to CFU with calibration curve and record in representative table below. | ||
Treatment 1: M9, 4°C | '''Treatment 1:''' ''M9, 4°C'' | ||
Blank OD = | |||
'''Blank OD =''' | |||
{| style="background:#F5FAFF; border: 1px solid #aabadd; color:Black" | {| style="background:#F5FAFF; border: 1px solid #aabadd; color:Black" | ||
|- style="background:#aabadd; color:Black" | |- style="background:#aabadd; color:Black" | ||
| '''Time Point''' || Visible Observation || OD<sub>600</sub> (abs units) || Number of Cells(CFU/ml)|| | | '''Time Point''' || '''Visible Observation''' || '''OD<sub>600</sub> (abs units)''' || |'''Number of Cells(CFU/ml)'''|| | ||
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Revision as of 07:56, 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) | |
| 1 | ||||
| 2 | ||||
| 3 | ||||
| 4 | ||||
| 5 |
