When cultures reach OD600 = 0.7, split into 10 ml for biomass evaluation . #####%%%%% output "biomass evaluation" are not used
{"action": "split", "output": "biomass evaluation", "volume": ["10 ml"]}
Resuspend cells in 1 ml 50 mM sodium phosphate buffer with 24 mM TTC, incubate at 30 °C for 1 h on a rotary shaker (200 rpm).
{"action": "incubate", "output": "", "temperature": ["30 \u00b0C"], "time": ["1 h"], "device": ["rotary shaker (200 rpm)"]}
Centrifuge for 1 min at 13,000 x g, collect supernatants. #####%%%%% output "supernatants" are not used
{"action": "centrifuge", "output": "", "time": ["1 min"], "speed": ["13,000 x g"]}
Prepare formazan standard solutions in 99.5% DMSO, use 99.5% DMSO as blank.
{"action": "prepare", "output": "", "reagent": ["formazan standard solutions"]}
read absorbance at 510 nm, #####%%%%% output "formazan standards" are used in line12
{"action": "read", "output": "formazan standards", "length": ["510 nm"]}
Plot calibration curve of formazan standards, determine formazan amount from reactions, . #####%%%%% output "formazan amount" are not used
{"action": "determine", "output": "formazan amount", "reagent": ["formazan standards"]}
normalize data to cell biomass,
{"action": "normalize", "output": ""}
determine formazan amount per g cells
{"action": "determine", "output": ""}