Team:Tsinghua/Notebook/24 August 2010

From 2010.igem.org

Module I, Group 2c


Liagtion for PEMK 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

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


Ligation for PC

Chlr                 5ul
PSB1C13              3ul
T4 ligase(NEB)       1ul
T4 ligation Buffer   2ul
H2O                  9ul

Negative control

PSB1C13              3ul
T4 ligase(NEB)       1ul
T4 ligation Buffer   2ul
H2O                  14ul

Incubate at 16°C overnight.