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Team:Warsaw/Calendar-Main/15 May 2009
From 2009.igem.org
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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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