IGEM:IMPERIAL/2007/Projects/Cell by date/Implementation/Calibrating Absorbance: Difference between revisions

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====Reagent====
====Reagent====
 
#Experimental Sample
 
 


===Protocol===
===Protocol===

Revision as of 08:00, 2 August 2007

Calibrating Absorbance

Summary

Materials Required

Equipment

  1. Spectrophotometer set to wavelength 600nm
  2. Cuvettes


Reagent

  1. Experimental Sample

Protocol

  1. 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.
  2. Next, take absorbance readings at a wavelength of 600nm for each time point, and record the relative turbidity of the broth.
  3. Subtract the 'blank' to measure actual OD600 of culture.
  4. 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