Normalize RNA samples to 1 ng/μl.
{"action": "normalize", "output": "", "reagent": ["RNA samples"]}
Thaw qPCR kit components on ice, mix gently, . #####%%%%% output "controls" are used in line6
{"action": "thaw", "output": "", "reagent": ["qPCR kit components"], "temperature": ["ice"]}
Calculate the number of samples, including controls, . #####%%%%% output "template/" are used in line8
{"action": "calculate", "output": "template/", "reagent": ["controls"]}
set up duplicate PCR reactions for each template/assay combination
{"action": "set", "output": "", "reagent": ["template/"]}
Seal the wells, mix, briefly, : 50°C (2 min), 95°C (10 min), 95°C (15 sec), 60°C (1 min), for 40 cycles.
{"action": "seal", "output": "", "container": ["the wells"], "reagent": [""]}
run the following program #####%%%%% output "the following program" are used in line14
{"action": "run", "output": "the following program", "concentration": [""]}
Analyze data . #####%%%%% output "data" are not used
{"action": "evaluate", "output": "data", "reagent": ["the following program"]}
adjust conditions if results are inconsistent before repeating the qPCR setup #####%%%%% output "the qPCR setup" are not used
{"action": "modify", "output": "the qPCR setup", "time": [""]}