Dandekar & Chandler:AHL Extraction: Difference between revisions
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# After the 10 minute rest, pipette off the clear acetate layer only being careful to not draw up any of the bottom cell debris layer. | # After the 10 minute rest, 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. | # 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 step 4. | # Repeat steps 1-4 and add acetate supernatant to collection tube from step 4. | ||
# Place acetate filled tubes into N2 evaporator | # 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. | # Suspend dried signal in 100uL of ethyl acetate, screw sealable cap on tightly, wrap in parafilm and place in -20 freezer. | ||
Revision as of 14:26, 22 January 2014
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 10 minute rest, 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.
- 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.
