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Team:Tsinghua/Notebook/24 August 2010
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(Difference between revisions)
(New page: == Module I, Group 2b == After purification, ligation reaction took place in a mixed system as following. eGFP 1ul mCherry 2ul Kan 1ul T4...) |
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== Module I, Group 2b == | == Module I, Group 2b == | ||
| + | ---- | ||
| + | |||
| + | Liagtion for PEMK | ||
After purification, ligation reaction took place in a mixed system as following. | After purification, ligation reaction took place in a mixed system as following. | ||
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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. | 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. | ||
Revision as of 20:25, 26 October 2010
Module I, Group 2b
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.
