The modified protocol (http://openwetware.org/wiki/Dandekar & Chandler:AHL Extraction): Difference between revisions
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(New page: ==AHL signal extraction from casein== In a large test tube (18 mm) containing the sample you wish to test: # Add 5mL of ethyl acetate '''AND''' "a spatula scoopful of NaCl" and vortex on...) |
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Latest revision as of 12:57, 12 October 2016
AHL signal extraction from casein
In a large test tube (18 mm) containing the sample you wish to test:
- Add 5mL of ethyl acetate AND "a spatula scoopful of NaCl" and vortex on high for 30 sec.
- Centrifuge at room temp for 10 min to separate phases.
- Pipette off the clear acetate layer only being 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. This type of vial works well. (Fisher 14-962-26F)
- Repeat steps 1-4 and add acetate supernatant to collection tube from step 4.
- Place acetate filled tubes into N2 evaporator until all acetate has evaporated off. (Keep on low level to prevent splashing)
- Suspend dried signal in 100uL of ethyl acetate, screw sealable cap on tightly, wrap in parafilm and place in -20 freezer.
