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Team:Goettingen/week18-3
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| - | <h2><b> | + | <h2><b>V08_29 </b></h2><br> |
| - | <b> | + | <b>Inoculation of LB-media for qrtPCR of different promoter constructs</b><br> |
<ul> | <ul> | ||
| - | <li>Experiment: <br> | + | <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> | ||
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| + | <h2><b>V08_30 </b></h2><br> | ||
| + | <b>First Test for Isolaton of RNA, Synthesis of cDNA and first qrtPCR approach</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> | ||
| + | First approach of qrtPCR with different delutions of cDNA in H20<br> | ||
| + | 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> | ||
| + | cDNA which was 1:10 diluted in H2O showed the best results for qrtPCR approach of our promoter constructs. | ||
| + | </li> | ||
| + | |||
</ul> | </ul> | ||
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Revision as of 14:57, 24 September 2012
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#3 Chemoreceptor Library - 18th WeekBack to overview
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