Team:Goettingen/week18-3

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Latest revision as of 20:28, 26 September 2012


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#3 Chemoreceptor Library - 18th Week

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V08_29

Inoculation of LB-media for qrtPCR of different promoter constructs

Experiment:
5 ml of LB-media with 50 µg/mL chloramphenicol were inoculated with E. coli strain BL21 haboring the vector pSB1C3 with following combinations of promoters and Tar receptor constructs:

- Tar receptor under the control of constitutive promoter J23100
- Tar receptor under the control of constitutive promoter J23104
- Tar receptor under the control of constitutive promoter J23105
- Tar receptor under the control of constitutive promoter J23106
- Tar receptor under the control of constitutive promoter J23109
- Tar receptor under the control of constitutive promoter J23112
- Tar receptor under the control of constitutive promoter J23113
- Tar receptor under the control of constitutive promoter J23114

For more details click here.

Cultures were incubated overnight at 37 °C, shaking.

V08_30

First Test for Isolaton of RNA, Synthesis of cDNA and qrtPCR approach

Experiment:
RNA was isolated using Nuceleus Spin RNA II (Machery and Nagel).
cDNA synthesis was performed using Reverse Transcription Kit (Qiagen).
First approach of qrtPCR with different delutions of cDNA in H₂0
For a detailed protocol click here.
Observations & Results
cDNA which was 1:10 diluted in H₂0 showed the best results for qrtPCR approach of our promoter constructs.

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@@ Line 30: / Line 30: @@
 <ul>
 <li>Experiment: <br>
-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>
+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>
@@ Line 39: / Line 39: @@
 - 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>
+- Tar receptor under the control of constitutive promoter J23114<br><br>
 For more details <a href="https://2012.igem.org/Team:Goettingen/iGEM/Parts">click here</a>.<br>
 <br>
@@ Line 58: / Line 58: @@
 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>
+First approach of qrtPCR with different delutions of cDNA in H<sub>2</sub>0<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.
+cDNA which was 1:10 diluted in H<sub>2</sub>0 showed the best results for qrtPCR approach of our promoter constructs.
 </li>