Team:Warsaw/Calendar-Main/15 May 2009

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</li> </ul>

Results: <img src="http://2009.igem.org/wiki/images/c/c8/2009.05.15.jpg"/>  Gel (from left)</li> </ul> <ol> GeneRuler DNA Ladder Mix #SM0333 (Fermentas)</li> B1001 ?</li> C0012 OK</li>

</ol>

Discussion:  The B1001 brick is too small to show up on a 1,5% agarose gel and should be tested by direct sequencing.</li> The C0012 brick looks OK</li> </ul>