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

From OpenWetWare

(Difference between revisions)
Jump to: navigation, search
(New page: ==A<sub>420</sub> versus o-nitrophenol concentration== *Make up a solution of ~5 mL solution (640 &mu;L permeabilization solution, 4.8 mL substrate solution without ONPG and 160 &m...)
Line 1: Line 1:
==A<sub>420</sub> versus [[ONP|o-nitrophenol]] concentration==
==A<sub>420</sub> versus [[ONP|o-nitrophenol]] concentration==
-
*Make up a solution of ~5 mL solution (640 &mu;L permeabilization solution, 4.8 mL substrate solution without ONPG and 160 &mu;L EZ rich media supplemented with kanamycin and AHL).
+
#Make up a solution of 7 mL to control for background absorbance in &beta;-galactosidase assays
 +
#*800 &mu;L permeabilization solution
 +
#*6 mL substrate solution without ONPG
 +
#*200 &mu;L EZ rich media supplemented with kanamycin and AHL
 +
#Make up 1mL of 4 mg/mL ONP.
 +
#Add 350 &mu;L of 4mg/mL solution to the first well.
 +
#Move 175 &mu;L of previous well to next well.
 +
#Add 175 &mu;L of background solution to that well.
 +
#Repeat until the well's solution looks totally clear.

Revision as of 13:54, 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.
Personal tools