Team:Goettingen/week19-3

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<h2><b>VX_Y </b></h2><br>
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<h2><b>V09_03 </b></h2><br>
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<b>Titel</b><br>
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<b>Inoculation of LB-media for qrtPCR of different promoter constructs</b><br>
<ul>
<ul>
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<li>Experiment: <br>hier text reinschreiben</li>
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<li>Experiment: <br>
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5 ml of LB-media with 50 µg/ml chloramphenicol were inoculated with <i>E. coli</i> strain BL21 haboring the vector pSB1C3 with following combinations of promoters and Tar receptor constructs:<br>
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<br>
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- Tar receptor under the control of constitutive promoter J23100<br>
 +
- Tar receptor under the control of constitutive promoter J23104<br>
 +
- Tar receptor under the control of constitutive promoter J23105<br>
 +
- Tar receptor under the control of constitutive promoter J23106<br>
 +
- Tar receptor under the control of constitutive promoter J23109<br>
 +
- Tar receptor under the control of constitutive promoter J23112<br>
 +
- Tar receptor under the control of constitutive promoter J23113<br>
 +
- Tar receptor under the control of constitutive promoter J23114<br>
 +
For more details <a href="https://2012.igem.org/Team:Goettingen/iGEM/Parts">click here</a>.<br>
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<br>
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Cultures were incubated overnight at 37 °C, shaking.</li>
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<h2><b>V09_04_ </b></h2><br>
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<b>V09_04_1 Isolaton of RNA, Synthesis of cDNA and qrtPCR </b><br>
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<ul>
 +
<li>Experiment: <br>
 +
RNA was isolated using Nuceleus Spin RNA II (Machery and Nagel).<br>
 +
cDNA synthesis was performed using Reverse Transcription Kit (Qiagen).<br>
 +
qrtPCR with promoter constructs was performed, for a detailed protocol <a href="https://2012.igem.org/Team:Goettingen/Project/Methods#-.3E_Experimental_design">click here</a>.<br>
 +
</li>
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<li>Observations & Results<br>
 +
Strong variations of CP-values, so we obtained no useful data for calculation of expression rates. Experiment needs to be repeated. </li>
 +
</ul>
 +
<br>
 +
 +
<b>V09_04_2 Inoculation of LB-media for qrtPCR of different promoter constructs </b><br>
 +
<ul>
 +
<li>Experiment: <br>
 +
5 ml of LB-media with 50 µg/ml chloramphenicol were inoculated with <i>E. coli</i> strain BL21 haboring the vector pSB1C3 with following combinations of promoters and Tar receptor constructs:<br>
 +
<br>
 +
- Tar receptor under the control of constitutive promoter J23100<br>
 +
- Tar receptor under the control of constitutive promoter J23104<br>
 +
- Tar receptor under the control of constitutive promoter J23105<br>
 +
- Tar receptor under the control of constitutive promoter J23106<br>
 +
- Tar receptor under the control of constitutive promoter J23109<br>
 +
- Tar receptor under the control of constitutive promoter J23112<br>
 +
- Tar receptor under the control of constitutive promoter J23113<br>
 +
- Tar receptor under the control of constitutive promoter J23114<br>
 +
For more details <a href="https://2012.igem.org/Team:Goettingen/iGEM/Parts">click here</a>.<br>
 +
<br>
 +
Cultures were incubated overnight at 37 °C, shaking.</li>
 +
 +
 +
<br></td></tr>
 +
</table>
 +
<br>
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<table cellpadding="20 px" border="1" bordercolor="black" valign="top">
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<h2><b>V09_05 </b></h2><br>
 +
<b>Isolaton of RNA, Synthesis of cDNA and qrtPCR</b><br>
 +
<ul>
 +
<li>Experiment: <br>
 +
RNA was isolated using Nuceleus Spin RNA II (Machery and Nagel).<br>
 +
cDNA synthesis was performed using Reverse Transcription Kit (Qiagen).<br>
 +
qrtPCR with promoter constructs was performed, for a detailed protocol <a href="https://2012.igem.org/Team:Goettingen/Project/Methods#-.3E_Experimental_design">click here</a>.<br>
 +
</li>
 +
<li>Observations & Results<br>
 +
Strong variations of CP-values, so we obtained no useful data for calculation of expression rates. Experiment needs to be repeated. </li>
 +
</ul>
 +
<br>
 +
<br></td></tr>
 +
</table>
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<br>
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Revision as of 15:34, 24 September 2012

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