Team:Goettingen/week7-2

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Revision as of 10:05, 14 September 2012

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V06_11_1 Miniprep of the new flhDC-promoter constructs

Experiment:
Minipreps were conducted with PeqGOLD MiniPrep Kit (Peqlab) according to the user manual.

V06_11_2 Chemical retransformartion of the new flhDC-promoter constructs into E. coli (DH10B)

Experiment:
For the chemical retransformation the standard protocol for transformation was followed. The following constructs were transformed into E. coli:
20G - #1
20G - #2
2G - #1
2G - #2
2G - #3
20I - #2
20I - #3
18O - #2
18O - #3
18K - #1
18K - #2
18K - #3
Observations & Results:
The retransformation was successful since all plates showed numeours colonoes except the negative control.

Preparation of over night cultures

Experiment:
In order to gain further plasmid material over night cultures were prepared for subsequent plasmid isolation.

Preparation of over night cultures

Experiment:
Over night cultures were preaped of three colonies of each constructs in order to isolate the plasmids.

V06_08_1 Miniprep of the new flhDC-promoter constructs

Experiment:
Minipreps were conducted with PeqGOLD MiniPrep Kit (Peqlab) according to the user manual.

V06_08_2 Test digestion of the new flhDC-promoter constructs

Experiment:
In order to prove correct insertion of flhDC a test digestion was performed using SpeI and XbaI according to the protocol.
Observations & Results:
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.

Important pages:
Home; Team; Official Team Profile; Project; Parts submitted to the Registry; Modeling; Notebook; Saftey; Attributions

@@ Line 542: / Line 542: @@
 <ul>
 <li>Experiment:  <br>
-For the chemical retransformation the standard protocol for transformation was followed. The following constructs were transformed into E. coli:
+For the chemical retransformation the standard protocol for transformation was followed. The following constructs were transformed into E. coli:<br>
 G - #1 <br>
 G - #2 <br>
@@ Line 566: / Line 566: @@
 <tr bordercolor="black" valign="top">
 <td width="900" bordercolor="black" valign="top">
-<h2><b>V06_06 </b></h2><br>
+<h2><b>V06_12 </b></h2><br>
-<b>Chemical transformartion of the <i>flhDC</i>-promoter constructs into <i>E. coli</i> (DH10B)</i></b><br>
+<b>Preparation of over night cultures</i></b><br>
 <ul>
 <li>Experiment: <br>
-For the chemical transformation the standard protocol was followed.
+In order to gain further plasmid material over night cultures were prepared for subsequent plasmid isolation.
-<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.
-</ul>
 <br></td></tr>
 </table>