Talk:Knight:Beta-galactosidase assay/96 well format: Difference between revisions
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(New page: ==A<sub>420</sub> versus o-nitrophenol concentration== *Make up a solution of ~5 mL solution (640 μL permeabilization solution, 4.8 mL substrate solution without ONPG and 160 &m...) |
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==A<sub>420</sub> versus [[ONP|o-nitrophenol]] concentration== | ==A<sub>420</sub> versus [[ONP|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. | |||
Revision as of 11:54, 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.
