**** Due to the coolant leak in the level1 cold room, please be advised that the Golden-gate order collection box will now be located in the level2 cold room (room #231) until further notice. ****
This page contains the recipe and reaction condition for assembling multiple overlapping fragments.
Read this article for an overview of Gibson Assembly and primer design.
5X ISO buffer. This recipe makes 6ml.
Gibson Assembly Enzyme Mix. This recipe makes 1.2ml
1. Thaw a 15 μl aliquot of Gibson Assembly Enzyme Mix on ice.
2. Add 5 μl of DNA (equimolar amounts of each fragment to a total of 50-200 ng)
3. Mix gently.
5. Incubate at 50 °C for 15 to 60 min.
6. Transform 1 – 5 μL.