Team:Goettingen/week7-2

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#2 Speed Improvement - 7th week

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V06_1

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.

V06_06

Chemical transformartion of the flhDC-promoter constructs into E. coli (DH10B)

Experiment:
For the chemical transformation the standard protocol was followed.
Observations & Results:
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.

V06_07

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

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.

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@@ Line 525: / Line 525: @@
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-<table cellpadding="20 px" border="1" bordercolor="black" valign="top">
-<tr bordercolor="black" valign="top">
-<td width="900" bordercolor="black" valign="top">
-<h2><b>V06_04 </b></h2><br>
-<b>Chemical retransformartion of the <i>flhDC</i>-promoter constructs into <i>E. coli</i> (DH10B)</i></b><br>
-<ul>
-<li>Experiment: <br>
-For the chemical retransformation the standard protocol for transformation was followed. The following constructs were transformed into
-E. coli: <br>
-E - #2 <br>
-E - #3 <br>
-M - #2 <br>
-M - #3 <br>
-C - #1 <br>
-C - #2 <br></li>
-<li>Observations & Results: <br>
-The retransformation was successful since all plates showed numeours colonoes except the negative control.
-<br> </li>
-</ul>
-<br></td></tr>
-</table>
 <br>
 <p align="justify" style="line-height:1.6em">
@@ Line 553: / Line 531: @@
 <tr bordercolor="black" valign="top">
 <td width="900" bordercolor="black" valign="top">
-<h2><b>V06_05 </b></h2><br>
+<h2><b>V06_1 </b></h2><br>
-<b>V06_05_1 Preparative double digestion of <i>flhDC</i> and plasmids containing further promoter</b><br>
+<b>V06_11_1 Miniprep of the new <i>flhDC</i>-promoter constructs</b><br>
 <ul>
 <li>Experiment:  <br>
-In order to clone <i>flhDC</i> behind further promoters, the purified PCR product was digested with XbaI and PstI, whereas the BioBrick plasmids were cut with SpeI and PstI. The digestion was performed as described in the protocol. This time the follwing promoters were used: <br>
+Minipreps were conducted with PeqGOLD MiniPrep Kit (Peqlab) according to the user manual.
-J23113 - 20G - x21 <br>
-J23109  - 2G - x106 <br>
-J23114 - 20I - x256 <br>
-J23106 - 18O - x1185 <br>
-J23104 - 18K - x 1831 <br>
 </ul>
 <br>
-<b>V06_05_2  Insertion of <i>flhDC</i> into the BioBrick plamids</b><br>
+<b>V06_11_2 Chemical retransformartion of the new flhDC-promoter constructs into E. coli (DH10B)</b><br>
 <ul>
 <li>Experiment:  <br>
-The ligation of the digested flhDC with the BioBrick plasmids was conducted as described in the protocol. <br> </li>
+For the chemical retransformation the standard protocol for transformation was followed. The following constructs were transformed into E. coli:
+G - #1 <br>
+G - #2 <br>
+G - #1 <br>
+G - #2 <br>
+G - #3 <br>
+I - #2 <br>
+I - #3 <br>
+O - #2 <br>
+O - #3 <br>
+K - #1 <br>
+K - #2 <br>
+K - #3 <br></li>
+<li>Observations & Results: <br>
+The retransformation was successful since all plates showed numeours colonoes except the negative control.
 </ul>
 <br></td></tr>