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Team:Warsaw/Calendar-Main/22 August 2009
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<p>Task 1:</p> | <p>Task 1:</p> | ||
<ul> | <ul> | ||
| - | <li>Electrophoresis of plasmid isolated and subsequently digested in: <a href="https://2009.igem.org/Team:Warsaw/Calendar-Main/ | + | <li>Electrophoresis of plasmid isolated and subsequently digested in: <a href="https://2009.igem.org/Team:Warsaw/Calendar-Main/21_August_2009">21.08.09</a></li></ul> |
<p>Results:</p> | <p>Results:</p> | ||
| - | <p>All samples of cro CDS on <a href="http://partsregistry.org/Part:pSB1A3"><span style="color: black">pSB1A3</a></span> are without insert</p> | + | <center><img src="https://static.igem.org/mediawiki/2009/c/c5/P53%2BRBS_cro%2BRBS_22_08_09.png" width="70%" heigth="70%"></center> |
| + | <font face="Times New Roman" size="3"><p><div style="text-align: center;">All samples of cro CDS on <a href="http://partsregistry.org/Part:pSB1A3"><span style="color: black">pSB1A3</a></span> are without insert (samples 1-8)<br/>All ligations of p53 CDS were correct (samples A-D)</div></p></font> | ||
<p><b>Comment:</p></b> | <p><b>Comment:</p></b> | ||
<p>The most probable explanation of this situation is circularization of empty plasmid because I have forgotten to dephosphorylate vector using CIAP</p> | <p>The most probable explanation of this situation is circularization of empty plasmid because I have forgotten to dephosphorylate vector using CIAP</p> | ||
| + | <br/> | ||
<p>Task 2:</p><ul><li>Prepare the bacterial cultures for isolation of following construct: <a href="http://partsregistry.org/Part:BBa_C0040"><span style="color: black">BBa_C0040</a></span> with <a href="http://partsregistry.org/Part:BBa_B0032"><span style="color: black">BBa_B0032</a></span> + <a href="http://partsregistry.org/Part:BBa_R0080"><span style="color: black">BBa_R0080</a></span> on <a href="http://partsregistry.org/Part:pSB1A3"><span style="color: black">pSB1A3</a></span> plasmid</li></ul> | <p>Task 2:</p><ul><li>Prepare the bacterial cultures for isolation of following construct: <a href="http://partsregistry.org/Part:BBa_C0040"><span style="color: black">BBa_C0040</a></span> with <a href="http://partsregistry.org/Part:BBa_B0032"><span style="color: black">BBa_B0032</a></span> + <a href="http://partsregistry.org/Part:BBa_R0080"><span style="color: black">BBa_R0080</a></span> on <a href="http://partsregistry.org/Part:pSB1A3"><span style="color: black">pSB1A3</a></span> plasmid</li></ul> | ||
| + | <h3><div style="text-align: center;">Making BioBricks' stocks</div></h3> | ||
| + | <h4>Justyna</h4> | ||
| + | <br/> | ||
| + | <p>Task:</p> | ||
| + | <ul> | ||
| + | <li>Making the glycerol stocks of bacteria bearing BioBricks.</li></ul> | ||
| + | <br><br> | ||
| + | <li>Following stocks were prepared: | ||
| + | <br>B0032+C0040, C0051+B0032, E0032+B0032, E0022+BOO32, P53+pSB1A3.</li> | ||
| + | <BR> | ||
| - | + | <h3><div style="text-align: center;">Making of the plac-RBS-llo-intA part</div></h3> | |
| + | <h4>Jarek</h4> | ||
| + | <br /> | ||
| + | <p>Tasks:</p> | ||
| + | <ul> | ||
| + | <li>Digestion of the the plac+RBS+llo and internalin A with PstI endonuclease.</li> | ||
| + | <li>Separation of digestion products in 0,8% agarose gel.</li> | ||
| + | <li>Cuting the agarose blocks containing the digested DNA from gel.</li> | ||
| + | <li>Preparation of pure DNA samples from agarose blocks with A&A GelOut kit.</li> | ||
| + | <li>Preparation of ligation containing digested internalin A and vector containing plac+RBS+llo part.</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> | </html> | ||
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| + | |||
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{{WarNotebookEnd}} | {{WarNotebookEnd}} | ||
Latest revision as of 05:19, 14 September 2009
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Assembly of endosomal detection operon
Marcin
Task 1:
- Electrophoresis of plasmid isolated and subsequently digested in: 21.08.09
Results:
All samples of cro CDS on pSB1A3 are without insert (samples 1-8)
All ligations of p53 CDS were correct (samples A-D)
All ligations of p53 CDS were correct (samples A-D)
Comment:
The most probable explanation of this situation is circularization of empty plasmid because I have forgotten to dephosphorylate vector using CIAP
Task 2:
- Prepare the bacterial cultures for isolation of following construct: BBa_C0040 with BBa_B0032 + BBa_R0080 on pSB1A3 plasmid
Making BioBricks' stocks
Justyna
Task:
- Making the glycerol stocks of bacteria bearing BioBricks.
B0032+C0040, C0051+B0032, E0032+B0032, E0022+BOO32, P53+pSB1A3.
Making of the plac-RBS-llo-intA part
Jarek
Tasks:
- Digestion of the the plac+RBS+llo and internalin A with PstI endonuclease.
- Separation of digestion products in 0,8% agarose gel.
- Cuting the agarose blocks containing the digested DNA from gel.
- Preparation of pure DNA samples from agarose blocks with A&A GelOut kit.
- Preparation of ligation containing digested internalin A and vector containing plac+RBS+llo part.
Ania
Tasks:
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