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From 2010.igem.org

Module I, Group 2b

After purification, ligation reaction took place in a mixed system as following.

eGFP                 1ul
mCherry              2ul
Kan                  1ul
T4 ligase(NEB)       1ul
T4 ligation Buffer   2ul
H2O                  13ul

Negative control

T4 ligase(NEB)       1ul
T4 ligation Buffer   2ul
H2O                  17ul

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Incubate at 16°C. After 8h, add 2ul digested and dephophorylated PSB1Ca3 into reaction system. Continue to incubate at 16°C overnight.