Talk:Knight:Beta-galactosidase assay/96 well format

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#Add 175 μL of background solution to that well.
#Add 175 μL of background solution to that well.
#Repeat until the well's solution looks totally clear.
#Repeat until the well's solution looks totally clear.
 +
#Measure A<sub>420</sub> of each well in the plate reader.

Revision as of 13:55, 24 October 2007

A420 versus o-nitrophenol concentration

  1. Make up a solution of 7 mL to control for background absorbance in β-galactosidase assays
    • 800 μL permeabilization solution
    • 6 mL substrate solution without ONPG
    • 200 μL EZ rich media supplemented with kanamycin and AHL
  2. Make up 1mL of 4 mg/mL ONP.
  3. Add 350 μL of 4mg/mL solution to the first well.
  4. Move 175 μL of previous well to next well.
  5. Add 175 μL of background solution to that well.
  6. Repeat until the well's solution looks totally clear.
  7. Measure A420 of each well in the plate reader.
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