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Team:Warsaw/Calendar-Main/17 August 2009
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(New page: {{WarNotebook}} <!-- do not edit above me! --> <html> <h3><div style="text-align: center;">Assembly of endosomal detection operon</div></h3> <h4>Marcin</h4> <br/> <p>Task 1:</p> <ul> <li>...) |
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<p>Task 1:</p> | <p>Task 1:</p> | ||
<ul> | <ul> | ||
| - | <li> | + | <li>Isolation of <a href="http://partsregistry.org/Part:pSB1A3"><span style="color: black">pSB1A3</a></span> plasmids containing <a href="http://partsregistry.org/Part:BBa_B0032"><span style="color: black">BBa_B0032</a></span> and <a href="http://partsregistry.org/Part:BBa_E0022"><span style="color: black">BBa_E0022</a></span> </li></ul> |
<p>Methods:</p><ul> | <p>Methods:</p><ul> | ||
<li>Plasmids were isolated using the A&A plasmid mini kit. Detailed procedure of the isolation is described <a href="http://aabiot.com/products/dna_purification/plasmid_dna/plasmid_mini/protocol_plasmid_mini.pdf" here>here</a></li></ul><br/> | <li>Plasmids were isolated using the A&A plasmid mini kit. Detailed procedure of the isolation is described <a href="http://aabiot.com/products/dna_purification/plasmid_dna/plasmid_mini/protocol_plasmid_mini.pdf" here>here</a></li></ul><br/> | ||
| Line 23: | Line 23: | ||
16.5 μl MQ water</pre> | 16.5 μl MQ water</pre> | ||
</ul> | </ul> | ||
| + | <p>Results:</p> | ||
| + | <center><img src="https://static.igem.org/mediawiki/2009/3/3d/Digest_17_08_09.png" width="60%" heigth="60%"></center> | ||
<p><b>Comment:</p></b> | <p><b>Comment:</p></b> | ||
<p>All isolated plasmids had the expected construct.</p> | <p>All isolated plasmids had the expected construct.</p> | ||
| Line 50: | Line 52: | ||
16.5 μl MQ water</pre> | 16.5 μl MQ water</pre> | ||
</ul> | </ul> | ||
| + | <p>Results:</p> | ||
| + | <center><img src="https://static.igem.org/mediawiki/2009/1/1e/Isolation_17_08_09.png" width="60%" heigth="60%"></center> | ||
<p><b>Comment:</p></b> | <p><b>Comment:</p></b> | ||
| - | <p>None of isolated plasmids had the expected constructs. | + | <p>None of isolated plasmids had the expected constructs. Ligations must be prepared once again.</p> |
<br/> | <br/> | ||
<p>Task 6:</p> | <p>Task 6:</p> | ||
| Line 69: | Line 73: | ||
24 μl MQ water</pre> | 24 μl MQ water</pre> | ||
</ul> | </ul> | ||
| + | <p>Results:</p> | ||
| + | <center><img src="https://static.igem.org/mediawiki/2009/0/09/PSB_p53_R0080_C0040%2BRBS_digest_18_08_09.png" width="60%" heigth="65%"></center> | ||
</html> | </html> | ||
| + | |||
| + | <html> | ||
| + | <h3><div style="text-align: center;">Making of the plac-RBS-llo part</div></h3> | ||
| + | <h4>Jarek</h4> | ||
| + | <br /> | ||
| + | <p>Tasks:</p> | ||
| + | <ul> | ||
| + | <li>Transformating the competent ''E.coli'' strain TopF' with ligation (acquired from Marek) containing RBS-llo gene cloned into vector with plac promoter </li> | ||
| + | <li>Plating the transformated culure on LB+Amp plates</li> | ||
| + | </ul> | ||
| + | <br /> | ||
| + | |||
| + | <h3><div style="text-align: center;">Cloning switch 1 regulatory parts [ | ||
| + | <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K177012">K177012</a>-PcI.RBS.LacI, | ||
| + | <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K177033">K177033</a>-PcI.RBS.LacI.PcI.RBS.RFP.terminator, | ||
| + | <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K177011">K177011</a>-PLacI.RBS.cI.terminator, | ||
| + | <a href="http://partsregistry.org/Part:BBa_K177038">K177038</a>-PLacI.RBS.cI.terminator.PLacI.RBS.GFP.terminator | ||
| + | ] | ||
| + | into two compatible low copy number plasmids of different antibiotic resistance</div></h3> | ||
| + | <h4>Ania</h4> | ||
| + | <br/> | ||
| + | <p>Tasks:</p> | ||
| + | <ul> | ||
| + | <li> | ||
| + | </li> | ||
| + | </ul> | ||
| + | <br/> | ||
| + | |||
| + | </html> | ||
| + | |||
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{{WarNotebookEnd}} | {{WarNotebookEnd}} | ||
Latest revision as of 21:27, 13 September 2009
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Assembly of endosomal detection operon
Marcin
Task 1:
Methods:
- Plasmids were isolated using the A&A plasmid mini kit. Detailed procedure of the isolation is described here
Task 2:
- Digest previously isolated plasmids to verify the correctness of the ligation
Methods:
- Reaction mixture composition:
1 μl purified plasmid DNA product 0.5 μl XbaI (Fermentas) 0.5 μl PstI (Fermentas) 2 μl Buffer Tango (Fermentas) 16.5 μl MQ water
Results:
Comment:
All isolated plasmids had the expected construct.
Task 3:
- Prepare bacterial cultures to isolate following constructs:
Task 4:
- Isolate of following constructs:
Methods:
- Plasmids were isolated using the A&A plasmid mini kit. Detailed procedure of the isolation is described here
Task 5:
- Digest previously isolated plasmids to verify the correctness of the ligation
Methods:
- Reaction mixture composition:
1 μl purified plasmid DNA product 0.5 μl XbaI (Fermentas) 0.5 μl PstI (Fermentas) 2 μl Buffer Tango (Fermentas) 16.5 μl MQ water
Results:
Comment:
None of isolated plasmids had the expected constructs. Ligations must be prepared once again.
Task 6:
- Restriction digest of following constructs:
Methods:
- Reaction mixture composition:
20 μl purified plasmid DNA product 1 μl XbaI (Fermentas) or 0.5 SpeI (Fermentas) 1 μl PstI (Fermentas) 5 μl Buffer Tango (Fermentas) 24 μl MQ water
Results:
Making of the plac-RBS-llo part
Jarek
Tasks:
- Transformating the competent ''E.coli'' strain TopF' with ligation (acquired from Marek) containing RBS-llo gene cloned into vector with plac promoter
- Plating the transformated culure on LB+Amp plates
Ania
Tasks:
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