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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 51: | Line 51: | ||
</ul> | </ul> | ||
<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> | ||
Revision as of 11:50, 19 August 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
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
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
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