Team:Warsaw/Calendar-Main/30 September 2009

Bistable switch testing

Monika

Tasks:


 * Transformation of chemocompetent cells (Top10) with plasmid containing bistable switch

Methods:
 * Transformation of chemocompetent cells with 10&mu;l 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:  BBa_0032 and BBa_C0080 digestion 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</a>+BBa_R0080 part</a> and BBa_E0032</a> digestion</li></ul> Overnight digestion in 37°C, volume of 30µl Comment: Enzymes: BBa_C0040+BBa_R0080 part: SpeI + PstI BBa_E0032 part: XbaI + PstI Results:</P> above. Task 2:  Ligation of YFP gene and BBa_C0040+BBa_R0080</li></ul> 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</li> </ul>

Methods:  21 of the colonies were selected and transferred to a new plate.</li> Liquid cultures were established for plasmid isolation.</li> </ul>