Talk:Knight:Beta-galactosidase assay

500mM dibasic sodium phosphate

7.098g Na₂HPO₄ in 100mL H₂O

1% hexadecyltrimethylammonium bromide (CTAB)

100mg CTAB in 10mL H₂O

1% sodium deoxycholate

10mg sodium deoxycholate in 1mL H₂O

Light-sensitive. Store at 4°C.

1M sodium carbonate

• 52.995 g Na₂CO₃ in 500mL H₂O

Measuring repression as a function of A₆₀₀

1. Grow an overnight culture of P20060.E0433
2. Dilute back to A₆₀₀ = 0.001 (as measured on the Nanodrop) in two tubes (1mL each)
3. Let grow for 1 hour
4. Prepare the permeabilization solution and aliquot out 80 μL per microfuge tube.
5. Add 20 μL of 50 mM IPTG to both tubes and 1 μL AHL to one tube.
6. Every hour, measure the A₆₀₀ and add 20 μL to 3 microfuge tubes.
7. Proceed with the Knight:Beta-galactosidase assay as normal.
8. Plot the Miller Units as a function of A₆₀₀.