Ligation is a method of combining several DNA fragments into a single plasmid. This is often the step following a PCR (and a PCR cleanup) or a gel extraction. You can also do a "dirty" ligation, where you follow a certain number of digestions directly by a ligation.

1. Download the following spreadsheet : File:Team-EPF-Lausanne Ligation.xls
2. Fill in the pink areas with the vector and fragment concentration, their size and the ratio.
3. Add all the suggested ingredients order in a microcentrifuge tube, in the order they appear.
4. Ligate for 2 hours at 14ºC.
5. Immediately transform competent bacteria with the ligation product.

Note: This protocol hasn't been optimized for blunt-end ligation (though it might still work).