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Team:Warsaw/Calendar-Main/15 May 2009
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(New page: {{WarNotebook}} <!-- do not edit above me! --> <html> <br/> <h3>Testing of Biobrick parts obtained from the registry</h3> <h4><font color="black">Kamil</font></h4> <br /> <p>Tasks:</p> <...) |
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(100ug/ml) and kanamycin (30ug/ml)</li> | (100ug/ml) and kanamycin (30ug/ml)</li> | ||
<li>The plasmids were extracted using the Plasmid Mini kit (A&A Biotechnology).</li> | <li>The plasmids were extracted using the Plasmid Mini kit (A&A Biotechnology).</li> | ||
| - | <li><p> | + | <li><p>Digest mixture's composition:</p> |
| - | + | <pre>2ul Orange buffer (Fermentas) | |
| - | plasmid extract | + | 1ul EcoRI enzyme |
| - | <li>The | + | 1ul PstI enzyme |
| + | 3ul plasmid extract</pre> | ||
| + | The solution was topped up with H2O to 20ul. </li> | ||
| + | <li>The digest was kept at 37 °C for 3h. | ||
<li>The electrophoretic separation was carried out on 1,5% agarose gel</li> | <li>The electrophoretic separation was carried out on 1,5% agarose gel</li> | ||
</ul> | </ul> | ||
Latest revision as of 17:46, 17 September 2009
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Testing of Biobrick parts obtained from the registry
Kamil
Tasks:
- Amplify the plasmid in bacterial culture
- Extract plasmids
- Digest the extracted plasmids
Methods:
- Bacteria from a single colony were cultivated overnight (~12h) in 50ml of LB medium supplemented with ampicillin (100ug/ml) and kanamycin (30ug/ml)
- The plasmids were extracted using the Plasmid Mini kit (A&A Biotechnology).
Digest mixture's composition:
2ul Orange buffer (Fermentas) 1ul EcoRI enzyme 1ul PstI enzyme 3ul plasmid extract
The solution was topped up with H2O to 20ul.- The digest was kept at 37 °C for 3h.
- The electrophoretic separation was carried out on 1,5% agarose gel
Results:
- Gel (from left)
- GeneRuler DNA Ladder Mix #SM0333 (Fermentas)
- B1001 ?
- C0012 OK
Discussion:
- The B1001 brick is too small to show up on a 1,5% agarose gel and should be tested by direct sequencing.
- The C0012 brick looks OK
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