Talk:Knight:Beta-galactosidase assay/96 well format: Difference between revisions
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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 11:55, 24 October 2007
A420 versus o-nitrophenol concentration
- 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
- Make up 1mL of 4 mg/mL ONP.
- Add 350 μL of 4mg/mL solution to the first well.
- Move 175 μL of previous well to next well.
- Add 175 μL of background solution to that well.
- Repeat until the well's solution looks totally clear.
- Measure A420 of each well in the plate reader.