Team:Lyon-INSA/notebook

From 2012.igem.org

(Difference between revisions)
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<titre>For all purposes</titre>
<titre>For all purposes</titre>
<description>
<description>
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<ul>
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<i>Bacillus subtilis</i> transformation with the 2 shuttle vectors pBK35 and pBK36.
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<li>Bacillus subtilis transformation with the 2 shuttle vectors pBK35 and pBK36.</li>
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</ul>
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</description>
</description>
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<titre>For all purposes</titre>
<titre>For all purposes</titre>
<description>
<description>
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<ul>
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Screening of 12 clones per <i>Bacillus</i> transformation.
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<li>Screening of 12 clones per Bacillus transformation;</li>
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</ul>
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</description>
</description>
<titre>Surfactant</titre>
<titre>Surfactant</titre>
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<titre>For all purposes</titre>
<titre>For all purposes</titre>
<description>
<description>
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<ul>
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Erythromycin at different concentrations was spread on LB plates with in order to test the Erythromycin resistance of the <i>B. Subtilis</i> strain transformed with the shuttle vectors.
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<li>LB plates were spread  with Erythromycin at different concentration in order to test the Erythromycin resistance of the <i>B. Subtilis</i> strain transformed with the shuttle vectors</li>
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</ul>
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</description>
</description>
<titre>Surfactant</titre>
<titre>Surfactant</titre>
<description>
<description>
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<li>Unfortunately the screened clones (containing the ligations pBK42+pBK35 and pBK42+pBK36) did not have the right plasmid. Another ligation of the construction [P<sub>xyl</sub>-<i>sfp</i>-<i>arbB</i>-<i>lacI</i>] is attempted, this time after gel extraction and purification of the digested pBK42 plasmid with EcoRI and PstI. This way, the backbone is eliminated and the chances of ligating the construction instead of the plasmid are higher.</li>
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Unfortunately the screened clones (containing the ligations pBK42+pBK35 and pBK42+pBK36) did not have the right plasmid. Another ligation of the construction [P<sub>xyl</sub>-<i>sfp</i>-<i>arbB</i>-<i>lacI</i>] is attempted, this time after gel extraction and purification of the digested pBK42 plasmid with EcoRI and PstI. This way, the backbone is eliminated and the chances of ligating the construction instead of the plasmid are higher.
</description>
</description>
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titre>For all purposes</titre>
titre>For all purposes</titre>
<description>
<description>
-
<ul>
+
Multiple tubes containing LB medium supplemented with Erythromycin at different concentrations are inoculated with the <i>B. subtilis</i> strains containing the shuttle vectors pBK35 and pBK36 in order to test the antibiotic resistance. For the same purpose LB + Ery plates are spotted with the same cultures at different dilution ratios.
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<li>Multiple tubes containing LB medium supplemented with Erythromycin at different concentrations are inoculated with the <i>B. subtilis</i> strains containing the shuttle vectors pBK35 and pBK36 in order to test the antibiotic resistance. For the same purpose LB + Ery plates are spotted with the same cultures at different dilution ratios.</li>
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</ul>
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</description>
</description>
<titre>Surfactant</titre>
<titre>Surfactant</titre>
<description>
<description>
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<li>30 clones containing the construction [P<sub>xyl</sub>-<i>sfp</i>-<i>arbB</i>-<i>lacI</i>] in the shuttle vector are screened
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30 clones containing the construction [P<sub>xyl</sub>-<i>sfp</i>-<i>arbB</i>-<i>lacI</i>] in the shuttle vector are screened.
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</li>
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</description>
</description>
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<titre>For all purposes</titre>
<titre>For all purposes</titre>
<description>
<description>
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<ul>
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The strains that were supposed to contain the shuttle vectors did not grow, so we suspect that the clones are in fact mutants.
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<li>The strains that were supposed to contain the shuttle vectors did not grow, so we suspect that the clones are in fact mutants</li>
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</ul>
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</description>
</description>
<titre>Surfactant</titre>
<titre>Surfactant</titre>
<description>
<description>
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<li> Miniprep of the transformed clones with the construction [P<sub>xyl</sub>-<i>sfp</i>-<i>arbB</i>-<i>lacI</i>]. The gel electrophoresis shows that only two of them have the expected fragments.  
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Miniprep of the transformed clones with the construction [P<sub>xyl</sub>-<i>sfp</i>-<i>arbB</i>-<i>lacI</i>]. The gel electrophoresis shows that only two of them have the expected fragments.
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</li>
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</description>
</description>
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<titre>For all purposes</titre>
<titre>For all purposes</titre>
<description>
<description>
-
<ul>
+
<i>Bacillus subtilis</i> transformation with the shuttle vectors pBK35 and pBK36.
-
 
+
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<li><i>Bacillus subtilis</i> transformation with the shuttle vectors pBK35 and pBK36</li>
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</ul>
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</description>
</description>
<titre>Surfactant</titre>
<titre>Surfactant</titre>
<description>
<description>
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<li> Transformation of <i>B. subtilis</i> 168 and <i>B. subtilis abrB</i> strains with the construction [P<sub>xyl</sub>-<i>sfp</i>-<i>arbB</i>-<i>lacI</i>] in the pBK35 shuttle vector.  
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Transformation of <i>B. subtilis</i> 168 and <i>B. subtilis abrB</i> strains with the construction [P<sub>xyl</sub>-<i>sfp</i>-<i>arbB</i>-<i>lacI</i>] in the pBK35 shuttle vector.  
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</li>
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</description>
</description>
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<titre>For all purposes</titre>
<titre>For all purposes</titre>
<description>
<description>
-
<ul>
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12 clones transformed with the shuttle vectors are screened.
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+
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<li>12 clones transformed with the shuttle vectors are screened</li>
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</ul>
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</description>
</description>
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<titre>Surfactant</titre>
<titre>Surfactant</titre>
<description>
<description>
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<li> 24 clones transformed with the construction [P<sub>xyl</sub>-<i>sfp</i>-<i>abrB</i>-<i>lacI</i>] are screened;
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24 clones transformed with the construction [P<sub>xyl</sub>-<i>sfp</i>-<i>abrB</i>-<i>lacI</i>] are screened.
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</li>
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</description>
</description>
</jour>
</jour>
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<titre>For all purposes</titre>
<titre>For all purposes</titre>
<description>
<description>
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<ul>
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Multiple tubes containing  2 mL liquid LB medium are supplemented with Erythromycin at various concentrations in order to test the antibiotic resistance of the two shuttle vectors. Also, LB+Ery plates are spotted with liquid cultures with the <i>B. subtilis </i> strains containing the shuttle vectors.
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+
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<li>Multiple tubes containing  2 mL liquid LB medium are supplemented with Erythromycin at various concentrations in order to test the antibiotic resistance of the two shuttle vectors. Also, LB+Ery plates are spotted with liquid cultures with the <i>B. subtilis </i> strains containing the shuttle vectors</li>
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</ul>
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</description>
</description>
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<titre>Surfactant</titre>
<titre>Surfactant</titre>
<description>
<description>
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<li> 6 of the 24 clones are put in liquid culture for further testing in LB medium supplemented with xylose at a concentration of 2%;
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6 of the 24 clones are put in liquid culture for further testing in LB medium supplemented with xylose at a concentration of 2%.
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</li>
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</description>
</description>
</jour>
</jour>
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<titre>For all purposes</titre>
<titre>For all purposes</titre>
<description>
<description>
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<ul>
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Both strains grew in all tubes. However, there is a difference concerning OD<sub>600</sub> between the two strains. The test is repeated (with antibiotic concentrations ranging from 0 to 1.5 mg/mL) in order to make sure there is a notable difference between the two strains. </li>
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<li> Both strains grew in all tubes. However, there is a difference concerning the OD between the two strains. The test is repeated (with antibiotic concentrations ranging from 0 to 1,5 mg/mL) in order to make sure there is a notable difference between the two strains. </li>
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</ul>
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</description>
</description>
<titre>Surfactant</titre>
<titre>Surfactant</titre>
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<titre>For all purposes</titre>
<titre>For all purposes</titre>
<description>
<description>
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<ul>
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The results of the antibiotic resistance in liquid culture show that the <i>B. subtilis</i> strain containing pBK35 is less resistant than the one containing pBK36.</li>
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<li>The results of the antibiotic resistance in liquid culture show that the <i>B. subtilis</i> strain containing pBK35 is less resistant than the one containing pBK36.</li>
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</ul>
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</description>
</description>
<titre>Surfactant</titre>
<titre>Surfactant</titre>
<description>
<description>
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<li> The emulsion test and the biofilm formation test confirm the presence of <i>sfp</i> and <i>abrB</i> genes;  
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The emulsion test and the biofilm formation test confirm the presence of <i>sfp</i> and <i>abrB</i> genes;  
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</li>
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</description>
</description>
</jour>
</jour>

Revision as of 03:19, 27 September 2012



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