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From 2010.igem.org

YX's part

Picked clones and screened for the positive ones.

Sent for sequencing.

Module I, Group 2c

To decrease the entropy of the reaction system, we decide to try a new ligation condition-1/10 system, which means all reacgents in the system is 10 percentage of normal quanlity and the total volume of reaction system is 10ul.

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

eGFP                 0.5ul
mCherry              0.8ul
Kan                  0.3ul
T4 ligase(NEB)       0.5ul
T4 ligation Buffer   1ul
H2O                  6.4ul

Negative control

T4 ligase(NEB)       0.5ul
T4 ligation Buffer   1ul
H2O                  8ul

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Incubate at 16°C. After 8h, add 0.5ul digested and dephophorylated PSB1Ca3 into reaction system. Continue to incubate at 16°C overnight.