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Team:Goettingen/week6-2
From 2012.igem.org
(Difference between revisions)
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<ul> | <ul> | ||
<li>Experiment: <br> | <li>Experiment: <br> | ||
| - | For the chemical retransformation the standard protocol for transformation was followed. The following constructs were transformed into | + | For the chemical retransformation the standard <a href="https://2012.igem.org/Team:Goettingen/Project/Methods">protocol</a> for transformation was followed. The following constructs were transformed into |
E. coli: <br> | E. coli: <br> | ||
20E - #2 <br> | 20E - #2 <br> | ||
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<ul> | <ul> | ||
<li>Experiment: <br> | <li>Experiment: <br> | ||
| - | In order to clone <i>flhDC</i> behind further promoters, the purified PCR product was digested with <i>XbaI</i> and <i>PstI</i>, whereas the BioBrick plasmids were cut with <i>SpeI</i> and <i>PstI</i>. The digestion was performed as described in the protocol. This time the follwing promoters were used: <br> | + | In order to clone <i>flhDC</i> behind further promoters, the purified PCR product was digested with <i>XbaI</i> and <i>PstI</i>, whereas the BioBrick plasmids were cut with <i>SpeI</i> and <i>PstI</i>. The digestion was performed as described in the <a href="https://2012.igem.org/Team:Goettingen/Project/Methods">protocol</a>l. This time the follwing promoters were used: <br> |
J23113 - 20G - x21 <br> | J23113 - 20G - x21 <br> | ||
J23109 - 2G - x106 <br> | J23109 - 2G - x106 <br> | ||
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<ul> | <ul> | ||
<li>Experiment: <br> | <li>Experiment: <br> | ||
| - | The ligation of the digested <i>flhDC</i> with the BioBrick plasmids was conducted as described in the protocol. <br> </li> | + | The ligation of the digested <i>flhDC</i> with the BioBrick plasmids was conducted as described in the <a href="https://2012.igem.org/Team:Goettingen/Project/Methods">protocol</a>. <br> </li> |
</ul> | </ul> | ||
<br></td></tr> | <br></td></tr> | ||
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<ul> | <ul> | ||
<li>Experiment: <br> | <li>Experiment: <br> | ||
| - | For the chemical transformation the standard protocol was followed. | + | For the chemical transformation the standard <a href="https://2012.igem.org/Team:Goettingen/Project/Methods">protocol</a> was followed. |
<li>Observations & Results: <br> | <li>Observations & Results: <br> | ||
The transformation was successful since all plates showed numeours colonoes except the negative control. However, on the plastes with 18K and 18O few red colonies could be observed. These were again rejected. | The transformation was successful since all plates showed numeours colonoes except the negative control. However, on the plastes with 18K and 18O few red colonies could be observed. These were again rejected. | ||
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<b>V06_08_2 Test digestion of the new <i>flhDC</i>-promoter constructs</b><br> | <b>V06_08_2 Test digestion of the new <i>flhDC</i>-promoter constructs</b><br> | ||
<li>Experiment: <br> | <li>Experiment: <br> | ||
| - | In order to prove correct insertion of <i>flhDC</i> a test digestion was performed using <i>SpeI</i> and <i>XbaI</i> according to the protocol.<br> | + | In order to prove correct insertion of <i>flhDC</i> a test digestion was performed using <i>SpeI</i> and <i>XbaI</i> according to the <a href="https://2012.igem.org/Team:Goettingen/Project/Methods">protocol</a>.<br> |
<li>Observations & Results: <br> | <li>Observations & Results: <br> | ||
For each construct two to three clones hosting the correctly inserted gene in the plasmid could be obtained. Those colonies still containing the rfp gene and were thus rejected.</li> | For each construct two to three clones hosting the correctly inserted gene in the plasmid could be obtained. Those colonies still containing the rfp gene and were thus rejected.</li> | ||
Revision as of 14:14, 18 September 2012
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