Team:Washington/Protocols/Gib Purif.

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=Gibson Purification=
=Gibson Purification=
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*Prepare this mixture on ice to prevent the reaction from beginning early
 
# Add 5 volumes of buffer PB to 1 volume of the Gibson product.
# Add 5 volumes of buffer PB to 1 volume of the Gibson product.
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#
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# Pour/Pipet the mixture into a spin column (pink).
 +
# Centrifuge the sample for ~ 1 minute. Discard the flow-through. 
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# Wash the sample by adding 750 μL buffer PE and centrifuge for ~ 1 minute.
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# Centrifuge the sample for ~ 1 minute. Discard the flow-through. 
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# To Elute the DNA, place the spin column in a clean 1.5 μL (labeled) microcentrifuge. Add 30 μL of buffer EB and '''let Stand for 1 minute!'''
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#Centrifuge the sample for ~ 1 minute.
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# Proceed to the Transformation (Electroporation) protocol.

Latest revision as of 19:40, 24 September 2011


Gibson Purification

  1. Add 5 volumes of buffer PB to 1 volume of the Gibson product.
  2. Pour/Pipet the mixture into a spin column (pink).
  3. Centrifuge the sample for ~ 1 minute. Discard the flow-through.
  4. Wash the sample by adding 750 μL buffer PE and centrifuge for ~ 1 minute.
  5. Centrifuge the sample for ~ 1 minute. Discard the flow-through.
  6. To Elute the DNA, place the spin column in a clean 1.5 μL (labeled) microcentrifuge. Add 30 μL of buffer EB and let Stand for 1 minute!
  7. Centrifuge the sample for ~ 1 minute.
  8. Proceed to the Transformation (Electroporation) protocol.
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