Table 2.

PCR cycling parameters for Golden Gate Cloning reaction where there are no internal BsaI/SapI sites within the insert fragment(s).

37°C 2 min.	25–50X
16°C 5 min.
80°C 10 min.
‡37°C 30 min.

^‡Before the start of this step, 1 μl of fresh BsaI needs to be added to the reaction mix.