{"action": "grow", "device": [""], "length": [""], "output": "", "reagent": ["YPD media"]}
{"action": "harvest", "container": ["50 ml tubes"], "device": ["centrifugation"], "output": "sterile water", "speed": ["1,000 x g"], "time": ["5 min"]}
{"action": "wash", "output": "", "reagent": ["sterile water"], "temperature": ["room temperature"], "time": [""]}
{"action": "resuspend", "output": "", "volume": ["50 ml culture"]}
{"action": "incubate", "output": "DTT", "temperature": ["30 \u00b0C"], "time": ["30 min"]}
{"action": "add", "output": "", "reagent": ["DTT"]}
{"action": "incubate", "container": ["the suspension(s)"], "output": "", "reagent": [""], "temperature": ["30 \u00b0"], "time": ["60 min"]}
{"action": "add", "output": "", "reagent": ["sterile pre-cooled water"], "volume": ["40 ml"]}
{"action": "centrifuge", "container": ["the suspensions"], "force": [""], "output": "", "time": [""]}
{"action": "resuspend", "container": ["cell pellets"], "output": "", "reagent": ["sterile water"], "temperature": ["4 \u00b0C"], "volume": ["25 ml"]}
{"action": "repeat", "output": "", "reagent": [""]}
{"action": "resuspend", "container": ["cell pellets"], "output": "", "reagent": [""], "volume": ["5 ml"]}
{"action": "repeat", "output": "", "reagent": [""]}
{"action": "resuspend", "concentration": ["pre-cooled 1 M sorbitol"], "output": "", "volume": ["550 \u00b5l"]}
{"action": "add", "output": "", "reagent": ["highly purified transforming DNA (1 to 3 \u00b5g)"], "volume": ["5 to 10 \u00b5l"]}
{"action": "incubate", "output": "YPD media", "temperature": ["ice"], "time": ["10 min"]}
{"action": "add", "container": ["each cuvette"], "output": "", "reagent": ["YPD media"]}
{"action": "transfer", "container": ["the suspension"], "output": ""}
{"action": "add", "output": "", "reagent": ["electrocompetent cell suspensions"], "volume": ["45 \u00b5l"]}
{"action": "incubate", "container": ["the plate"], "output": "", "temperature": ["ice"], "time": ["10 min"]}
{"action": "make", "length": [""], "output": ""}
{"action": "cover", "container": ["the plate"], "device": ["a sterile lid"], "output": "YPD media"}
{"action": "transfer", "container": ["the cell suspension"], "output": "", "reagent": ["YPD media"]}
{"action": "use", "output": "", "volume": [""]}
{"action": "centrifuge", "container": ["the tubes"], "force": ["1,000 x g"], "output": "sterile water", "time": ["5 min"]}
{"action": "remove", "output": "", "reagent": ["sterile water"]}