Team:Washington/Protocols/Gib Rxn

From 2011.igem.org

(Difference between revisions)
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# Add ~20-50 ng of purified Backbone DNA to the reaction tube.  
# Add ~20-50 ng of purified Backbone DNA to the reaction tube.  
# Add ~20-50 ng of purified Insert DNA to the reaction tube.
# Add ~20-50 ng of purified Insert DNA to the reaction tube.
-
# Fill the remaining tubes with distilled H20 to ensure a final volume of '''20 uL'''
+
# Fill the remaining tubes with ice cold, distilled H20 to ensure a final volume of '''20 uL'''
# After making sure all tubes are appropriately labeled, incubate all samples for ~1 hour @ 50oC.
# After making sure all tubes are appropriately labeled, incubate all samples for ~1 hour @ 50oC.

Revision as of 04:57, 14 September 2011


Gibson Cloning/Assembly

  • Prepare this mixture on ice to prevent the reaction from beginning early
  1. Add 15uL of Gibson MasterMix to each tube.
  2. Add ~20-50 ng of purified Backbone DNA to the reaction tube.
  3. Add ~20-50 ng of purified Insert DNA to the reaction tube.
  4. Fill the remaining tubes with ice cold, distilled H20 to ensure a final volume of 20 uL
  5. After making sure all tubes are appropriately labeled, incubate all samples for ~1 hour @ 50oC.