From 2011.igem.org

pGA vector Assay

• 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.