Transfer 200 µl of serum into a 1.5 ml microfuge tube. #####%%%%% output "buffer" are used in line4
{"action": "transfer", "output": "buffer", "reagent": ["serum"], "container": ["a 1.5 ml microfuge tube"]}
Add 200 µl of buffer XBP. #####%%%%% output "the sample/XBP mixture" are used in line6
{"action": "add", "output": "the sample/XBP mixture", "reagent": ["buffer"]}
Add the sample/XBP mixture onto the exoEasy spin column. #####%%%%% output "buffer XWP" are used in line8
{"action": "add", "output": "buffer XWP", "reagent": ["the sample/XBP mixture"]}
Add 10 ml of buffer XWP to the exoEasy spin column.
{"action": "add", "output": "", "reagent": ["buffer XWP"], "volume": ["10 ml"]}
Transfer the spin column to a fresh collection tube. #####%%%%% output "Qiazol" are used in line12
{"action": "transfer", "output": "Qiazol", "container": ["the spin column", "a fresh collection tube"], "volume": [""]}
Add 700 μl Qiazol to the membrane of the spin column.
{"action": "add", "output": "", "reagent": ["Qiazol"], "volume": ["700 μl"]}
Transfer the flow-though, which is the lysate, to a PLG tube.<sup>1</sup> 13. #####%%%%% output "the flow-though" are used in line34
{"action": "transfer", "output": "the flow-though", "container": ["a PLG tube.<sup>1</sup> 13"]}
Incubate at room temperature for 5 minutes.
{"action": "incubate", "output": "", "temperature": ["room temperature"], "time": ["5 minutes"]}
Add 90 µl chloroform.
{"action": "add", "output": "", "volume": ["90 \u00b5l chloroform"], "mass": [""]}
Shake vigorously for 15 seconds.
{"action": "shake", "output": "", "time": ["15 seconds"]}
Incubate for 3 min at room temperature.
{"action": "incubate", "output": "", "temperature": ["room temperature"], "time": ["3 min"]}
Transfer the upper aqueous phase to a new microcentrifuge tube.
{"action": "transfer", "output": "", "container": ["a new microcentrifuge tube"]}
Carefully measure the aqueous phase .
{"action": "measure", "output": "", "container": ["the aqueous phase"]}
add 2 volumes of 100% ethanol
{"action": "add", "output": "", "reagent": ["100% ethanol"]}
do not delay moving on to the next step

Assemble a MinElute spin column in a new collection tube.
{"action": "assemble", "output": "", "container": ["a MinElute spin column"], "reagent": [""]}
Load up to 700 µl of the mixture from step 5, including any precipitate that may have formed, onto the column.
{"action": "load", "output": "", "volume": ["up to 700"], "device": [""], "reagent": ["the flow-though"]}
Repeat steps 23-25 until entire sample has been loaded. #####%%%%% output "buffer RWT" are used in line38
{"action": "move", "output": "buffer RWT", "time": ["23-25"]}
Add 700 μl of buffer RWT to the RNeasy MinElute spin column.
{"action": "add", "output": "", "reagent": ["buffer RWT"], "volume": ["700 µl"]}
Discard the flow-through. #####%%%%% output "the flow-through" are used in line44
{"action": "remove", "output": "the flow-through", "volume": [""]}
Discard the flow-through. #####%%%%% output "the flow-through" are used in line44
{"action": "remove", "output": "the flow-through", "volume": [""]}
Discard the collection tube with the flow-through.<sup>4</sup> 36. #####%%%%% output "the flow-through.<sup>4</sup> 36" are used in line52
{"action": "remove", "output": "the flow-through.<sup>4</sup> 36", "container": ["the collection tube"], "volume": [""], "reagent": ["the flow-through"]}
Transfer the RNeasy MinElute spin column into a new 2 ml collection tube \(supplied).
{"action": "transfer", "output": "", "device": ["RNeasy MinElute"], "container": ["a new 2 ml collection tube (supplied)"], "volume": [""]}
Open the lid of the spin column, .
{"action": "open", "output": "", "container": ["the spin column"]}
centrifuge at full speed \(14,000 xg) for 5 min to dry the membrane.<sup>5</sup> 38
{"action": "centrifuge", "output": "", "speed": ["full speed (14,000 xg)"], "container": ["membrane.<sup>5</sup>"], "time": ["5 min"]}
Discard the collection tube with the flow-through.<sup>4</sup> 39. #####%%%%% output "the flow-through.<sup>4</sup> 39" are not used
{"action": "remove", "output": "the flow-through.<sup>4</sup> 39", "container": ["the collection tube"], "volume": [""], "reagent": ["the flow-through.<sup>4</sup> 36"]}
Transfer the RNeasy MinElute spin column into a new 1.5 ml collection tube \(supplied). #####%%%%% output "RNase-free water" are used in line56
{"action": "transfer", "output": "RNase-free water", "device": ["RNeasy MinElute"], "container": ["a new 1.5 ml collection tube (supplied)"], "volume": [""]}
Add 14 μl RNase-free water directly to the center of the spin column membrane.<sup>6</sup> 40.
{"action": "add", "output": "", "reagent": ["RNase-free water"], "volume": ["14 µl"]}