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From 2010.igem.org

Module I, group 2c

Since it seems as if no ligation product of three fragments appeared in the PCR identification result, we strongly doubt that there is something wrong with the DraIII cutting site, perhaps there lies mutaion site. To confirm our proposal, we decide to conduct the ligation sepratatively and then PCR is run.

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Ligation for EM

eGFP                 1ul
mCherry              2ul
T4 ligase(NEB)       1ul
T4 ligation Buffer   2ul
H2O                  14ul

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Ligation for MK

Kan                  1ul
mCherry              2ul
T4 ligase(NEB)       1ul
T4 ligation Buffer   2ul
H2O                  14ul

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Incubate at 16°C overnight.