Team:Hawaii/Procols/DNA ligation

Protocol

 * 1) Combine 4 ul of construct one with 4 ul of construct 2.
 * 2) Add 1 ul of nanopure H20.
 * 3) Add 10 ul of 2x quick ligation reaction Buffer. In order to avoid shearing the DNA, mix by resuspending very slowly.
 * 4) Add 1 ul of Quick T4 DNA ligase and mix thoroughly by resuspending very slowly.
 * 5) Incubate at room temperature for 5 minutes.
 * 6) Cool on ice then transform, or store at -20oC

Notes:
 * Adjust amount of DNA as necessary. For ligation of 2 pieces of DNA, use 1:1 ratio of parts. For ligation of insert to vector, use 3:1 ratio of insert to vector. ~180ng of DNA can be ligated using the above protocol.


 * Reference: Quick Ligation Kit from NEB.


 * The Endy lab recommends using ~10 ng vector in a reaction of with 6:1 ratio of insert to vector. Total DNA concentration should not exceed 100ng for maximum ligation efficiency.


 * Reference: OpenWetWare