Add 4 μl of 160 mM KMnO4 to radiolabeled DNA (40 ng, 5,000-10,000 cpm) in 40 μl total volume. #####%%%%% output "5,000-10,000 cpm" are used in line6
{"action": "add", "output": "5,000-10,000 cpm", "reagent": ["160 mM KMnO4"], "volume": ["40 \u03bcl"]}
Incubate for 2 min at 30 °C, stop reaction with 4.8 μl β-mercaptoethanol, add 5.3 μl 500 mM EDTA. #####%%%%% output "phenol/chloroform" are used in line6
{"action": "add", "output": "phenol/chloroform", "reagent": ["500 mM EDTA"], "volume": ["5.3 μl"]}
Extract with phenol/chloroform, precipitate with ethanol, , .
{"action": "precipitate", "output": "", "reagent": ["ethanol"]}
dissolve in 70 μl 10% piperidine,
{"action": "dissolve", "output": "", "volume": ["70 \u03bcl"]}
incubate at 90 °C for 30 min
{"action": "incubate", "output": "", "temperature": ["90 \u00b0C"], "time": ["30 min"]}
Lyophilize, dissolve in 30 μl water, lyophilize again, repeat.
{"action": "repeat", "output": "", "volume": ["30 \u03bcl water"]}
Dissolve in 50 μl water, μl glycogen, .
{"action": "dissolve", "output": "", "volume": ["50 \u03bcl water"]}
add 4 μl 300 mM NaOAc, 1 #####%%%%% output "ethanol" are used in line18
{"action": "add", "output": "ethanol", "volume": ["4 \u03bcl 300 mM NaOAc,"]}
precipitate with ethanol #####%%%%% output "pellets" are used in line20
{"action": "precipitate", "output": "pellets", "reagent": ["ethanol"]}
Wash pellets with 70% ethanol, dry, dissolve in 5 μl electrophoresis loading buffer. #####%%%%% output "non-labeled DNA" are used in line22
{"action": "rinse", "output": "non-labeled DNA", "reagent": ["pellets"]}
For non-labeled DNA, follow similar steps. #####%%%%% output "similar steps" are used in line28
{"action": "go", "output": "similar steps", "reagent": ["non-labeled DNA"]}
, add 5'-32P-labeled primer, #####%%%%% output "primer extension" are used in line26
{"action": "add", "output": "primer extension", "reagent": ["5'-32P-labeled primer"]}
perform primer extension, ,
{"action": "perform", "output": "", "reagent": ["primer extension"]}
separate on denaturing gel
{"action": "separate", "output": "", "device": ["denaturing gel"]}
visualize by autoradiography #####%%%%% output "autoradiography" are not used
{"action": "visualize", "output": "autoradiography", "reagent": [""]}