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