Dandekar & Chandler:AHL Extraction

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AHL Signal Extraction

In a large test tube containing the sample you wish to test:

• Add 5mL of ethyl acetate and vortex on high for 30seconds
• Let the sample sit for 10 minutes to allow the two phases to separate out.
• After the the 10 minute rest, pipette off the clear acetate layer only. Be careful to not draw up any of the bottom cell debris layer.
• Deposit the clear acetate layer in a 10mL glass tube with a sealable air tight cap.
• Repeat steps 1-4 and add acetate supernatant to tube from first cycle.
• Place acetate filled tubes into N2 evaporator on medium until all acetate has evaporated off.
• Suspend dried signal in 100uL of ethyl acetate, screw sealable cap on tightly, wrap in parafilm and place in -20 freezer.