Dandekar & Chandler:AHL Extraction

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(AHL Signal Extraction)
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In a large test tube containing the sample you wish to test:
In a large test tube containing the sample you wish to test:
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* Add 5mL of ethyl acetate and vortex on high for 30seconds
+
# 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.   
+
# Let the sample sit for 10 minutes to allow the two phases to separate out.   
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* 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.  
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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.  
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* Deposit the clear acetate layer in a 10mL glass tube with a sealable air tight cap.   
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# Deposit the clear acetate layer in a 10mL glass tube with a sealable air tight cap.   
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* Repeat steps 1-4 and add acetate supernatant to tube from first cycle.  
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# Repeat steps 1-4 and add acetate supernatant to tube from step 4.  
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* Place acetate filled tubes into N2 evaporator on medium until all acetate has evaporated off.  
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# Place acetate filled tubes into N2 evaporator on medium until all acetate has evaporated off.  
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* 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 18:42, 14 November 2013

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AHL Signal Extraction

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

  1. Add 5mL of ethyl acetate and vortex on high for 30seconds
  2. Let the sample sit for 10 minutes to allow the two phases to separate out.
  3. 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.
  4. Deposit the clear acetate layer in a 10mL glass tube with a sealable air tight cap.
  5. Repeat steps 1-4 and add acetate supernatant to tube from step 4.
  6. Place acetate filled tubes into N2 evaporator on medium until all acetate has evaporated off.
  7. Suspend dried signal in 100uL of ethyl acetate, screw sealable cap on tightly, wrap in parafilm and place in -20 freezer.
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