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Team:Tsinghua/Notebook/15 September 2010
From 2010.igem.org
(Difference between revisions)
(New page: 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...) |
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| + | == 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. | 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. | To confirm our proposal, we decide to conduct the ligation sepratatively and then PCR is run. | ||
| + | ---- | ||
| + | Ligation for EM | ||
| + | eGFP 1ul | ||
| + | mCherry 2ul | ||
| + | T4 ligase(NEB) 1ul | ||
| + | T4 ligation Buffer 2ul | ||
| + | H2O 14ul | ||
| + | ---- | ||
| + | Ligation for MK | ||
| + | Kan 1ul | ||
| + | mCherry 2ul | ||
| + | T4 ligase(NEB) 1ul | ||
| + | T4 ligation Buffer 2ul | ||
| + | H2O 14ul | ||
| + | ---- | ||
| + | Incubate at 16°C overnight. | ||
Latest revision as of 21:57, 26 October 2010
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.
Ligation for EM
eGFP 1ul mCherry 2ul T4 ligase(NEB) 1ul T4 ligation Buffer 2ul H2O 14ul
Ligation for MK
Kan 1ul mCherry 2ul T4 ligase(NEB) 1ul T4 ligation Buffer 2ul H2O 14ul
Incubate at 16°C overnight.
