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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
- Add 15uL of Gibson MasterMix to each 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.
- 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.
