Day 1: Grow bacteria overnight in 10 ml 3% TH medium at 37 °C, 200 rpm.
{"action": "grow", "output": "", "concentration": ["10 ml 3% TH medium"], "temperature": ["37 \u00b0C"]}
Centrifuge bacteria (10 min, 5,000 rpm), remove supernatant, . #####%%%%% output "supernatant" are used in line6
{"action": "remove", "output": "supernatant", "reagent": [""]}
wash pellet twice with 10 ml 50 mM Tris/HCl (pH 7.4)
{"action": "wash", "output": "", "time": [""], "reagent": ["supernatant"]}
Incubate 100 µl bacteria with 10 µl test agent for 60 sec at RT.
{"action": "incubate", "output": "", "time": ["60 sec"], "temperature": ["RT"]}
Add 100 µl citrate plasma, incubate 35 min at 37 °C, 200 rpm. #####%%%%% output "bacteria" are used in line12
{"action": "add", "output": "bacteria", "time": ["35 min"], "temperature": ["37 \u00b0C"], "reagent": [""]}
Spin down bacteria (6 min, 5,000 rpm), wash pellets once with 50 mM Tris/HCl (pH 7.4). #####%%%%% output "pellets" are not used
{"action": "wash", "output": "pellets", "reagent": ["bacteria"]}
Incubate 30-60 min at 37 °C, , transfer supernatants to 96 well plate. #####%%%%% output "supernatants" are not used
{"action": "transfer", "output": "supernatants", "container": ["96 well plate"]}
For assay without bacteria, follow similar steps with modifications as specified. #####%%%%% output "modifications" are not used
{"action": "go", "output": "modifications", "reagent": ["bacteria"]}