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Team:Warsaw/Calendar-Main/30 September 2009
From 2009.igem.org
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| + | <h3><div style="text-align: center;">Cloning of the cro-box into the pSB1A3 plasmid</div></h3> | ||
| + | <h4>Kamil</h4> | ||
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| + | <p>Tasks:</p> | ||
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| + | <li>Transformatnt selection</li> | ||
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| + | <br /> | ||
| + | <p>Methods:</p> | ||
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| + | <li>21 of the colonies were selected and transferred to a new plate.</li> | ||
| + | <li>Liquid cultures were established for plasmid isolation.</li> | ||
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Latest revision as of 18:12, 12 October 2009
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Bistable switch testing
Monika
Tasks:
- Transformation of chemocompetent cells (Top10) with plasmid containing bistable switch
Methods:
- Transformation of chemocompetent cells with 10μl of DNA solution - procedure destribed here
- Planting on LB-agar medium supplemented with tetacycline
Results:
Successful, many colonies that express GFP.
Cloning of AraC coding sequence
Justyna
Task 1:
Overnight digestion in 37°C, volume of 30µl
Comment:
Enzymes:
BBa_B0032 part: SpeI + PstI
BBa_C0080 part: XbaI + PstI
Results:
Task 2:
- Ligation of AraC gene and BBa_B0032
4 hour long ligation in room temperature, volume of 25µl
Ligation mix:
BBa_B0032 7µl (~49ng) AraC sequence 15.5µl (~70ng) buffer 2.5µl ligase 1µl (Fermentas)
Fusion of YFP coding sequence and TetR coding sequence
Justyna
Task 1:
- BBa_C0040+BBa_R0080 part and BBa_E0032 digestion
Overnight digestion in 37°C, volume of 30µl
Comment:
Enzymes:
BBa_C0040+BBa_R0080 part: SpeI + PstI
BBa_E0032 part: XbaI + PstI
Results:
above.Task 2:
- Ligation of YFP gene and BBa_C0040+BBa_R0080
4 hour long ligation in room temperature, volume of 20µl
Ligation mix:
BBa_C0040+BBa_R0080 7µl (~50ng) YFP sequence 9.5µl (~114ng) buffer 2.0µl ligase 1µl (Fermentas)
Cloning of the cro-box into the pSB1A3 plasmid
Kamil
Tasks:
- Transformatnt selection
Methods:
- 21 of the colonies were selected and transferred to a new plate.
- Liquid cultures were established for plasmid isolation.
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