Ligation

Goal

Protocol

1. Set up the water bath at 16°C.


2. In an eppendorf tube, introduce :
   1 µL of T4 DNA ligase
   2 µL of T4 DNA ligase buffer (10X)
   4 µL (50 ng) of DNA vector
   12 µL (50 ng) of DNA insert
   to 20 µL of water
   
   

   SAFETY AND USEFUL RECOMMANDATION

   Those products are not dangerous at first sight, but in case of spreading or contacts with any part of the body, it is recommended to wash this part and to dry it after. Use a different pipette tip for each sample. It is also important to close tubes after putting the different reagents. DNA is more sensitive to its environment while it is not circulared in a plasmid and introduced into a cell. Therefore, a spreading has very few chances to lead to any bad consequences.

   
   
3. Incubate at room temperature for 10 minutes.