Add 1 ml of Chloroform:Methanol:Isopropanol (2:1:1) to 200 µl of plasma .
{"action": "add", "output": "", "reagent": ["Chloroform:Methanol:Isopropanol (2:1:1)"], "container": ["plasma"]}
Centrifuge at 12,000 rcf for 10 minutes at 4°C, then discard the supernatant. #####%%%%% output "the supernatant" are used in line12
{"action": "discard", "output": "the supernatant", "container": [""]}
Add 800 µl TCA/Acetone (50%) .
{"action": "add", "output": "", "volume": ["800 µl"], "reagent": ["TCA/Acetone (50%)"], "device": [""]}
incubate overnight at -20°C
{"action": "incubate", "output": "", "temperature": ["-20\u00b0"]}
Centrifuge at 12,000 rcf for 10 minutes at 4°C, then discard the supernatant. #####%%%%% output "the supernatant" are used in line12
{"action": "discard", "output": "the supernatant", "container": [""]}
Add 40% Ammonium sulfate .
{"action": "add", "output": "", "reagent": ["40% Ammonium sulfate"]}
incubate on ice for 30 minutes
{"action": "incubate", "output": "", "temperature": ["ice"], "time": ["30 minutes"]}
Centrifuge at 12,000 rcf for 10 minutes, discard the supernatant, . #####%%%%% output "the supernatant" are not used
{"action": "remove", "output": "the supernatant", "container": [""]}
Clean the pellet with a Bio-Rad clean-up kit, . #####%%%%% output "the pellet" are used in line22
{"action": "clean", "output": "the pellet", "device": ["Bio-Rad clean-up kit"]}
dissolve in 150-250 µl Rehydration buffer,
{"action": "dissolve", "output": "", "volume": ["150-250 µl"], "reagent": ["Rehydration buffer"]}
resolve through 10% SDS-PAGE
{"action": "resolve", "output": "", "concentration": ["10%"], "container": ["SDS-PAGE"], "length": [""], "reagent": ["the pellet"]}