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Team:Warsaw/Calendar-Main/23 July 2009
From 2009.igem.org
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<li>Reaction mixture: 1ul pAcGFP1-Mito (5ng), 1ul dNTPs (2mM), 1ul L-primer (5pmol/ul), 1ul R-primer (5pmol/ul), 1ul Pfu buffer, 4,5ul ddH2o, 0,5ul Pfu DNA polymerase</li> | <li>Reaction mixture: 1ul pAcGFP1-Mito (5ng), 1ul dNTPs (2mM), 1ul L-primer (5pmol/ul), 1ul R-primer (5pmol/ul), 1ul Pfu buffer, 4,5ul ddH2o, 0,5ul Pfu DNA polymerase</li> | ||
<li>PCR program: 5min 95C, 1min 95C, 1min gradient 48-60C, 1min 72C, go to 2-nd step (30x), 5min 72C</li> | <li>PCR program: 5min 95C, 1min 95C, 1min gradient 48-60C, 1min 72C, go to 2-nd step (30x), 5min 72C</li> | ||
| + | </ul> | ||
</html> | </html> | ||
| + | <html> | ||
| + | <h3>Cloning of the mgtc promoter into the pKSII+ plasmid</h3> | ||
| + | <h4>Kamil</h4> | ||
| + | <br /> | ||
| + | <p>Tasks:</p> | ||
| + | <ul> | ||
| + | <li>Transformant selection</li> | ||
| + | </ul> | ||
| + | <br /> | ||
| + | <p>Methods:</p> | ||
| + | <ul> | ||
| + | <li>The colonies that remained white after 48h were picked up and transferred to a new petri dish.</li> | ||
| + | <li>Liquid cultures were established along the way for plasmid purification.</li> | ||
| + | <li>Both the dish and the liquid cultures were incubated at 37°C overnight.</li> | ||
| + | </ul> | ||
| + | <br /> | ||
| + | </html> | ||
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{{WarNotebookEnd}} | {{WarNotebookEnd}} | ||
Revision as of 12:17, 24 July 2009
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PCR of Cox mitochondrial export sequence
Sebastian
Task 1:
- PCR with gradient temperature of anniling
Methods:
- Reaction mixture: 1ul pAcGFP1-Mito (5ng), 1ul dNTPs (2mM), 1ul L-primer (5pmol/ul), 1ul R-primer (5pmol/ul), 1ul Pfu buffer, 4,5ul ddH2o, 0,5ul Pfu DNA polymerase
- PCR program: 5min 95C, 1min 95C, 1min gradient 48-60C, 1min 72C, go to 2-nd step (30x), 5min 72C
Cloning of the mgtc promoter into the pKSII+ plasmid
Kamil
Tasks:
- Transformant selection
Methods:
- The colonies that remained white after 48h were picked up and transferred to a new petri dish.
- Liquid cultures were established along the way for plasmid purification.
- Both the dish and the liquid cultures were incubated at 37°C overnight.
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