Team:Goettingen/week10-2

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#2 Speed Improvement - 10th week

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V07_02

V07_02_1 Preparative double digestion of 20E-flhDC, 2G-flhDC, 18O-flhDC and pSB1C3

Experiment:
In order to clone the flhDC-promoter constructs intro pSB1C3, all components were digested with EcoRI and PstI according to the protocol. The restrictio products were subsequently separated on a gel in order to verify the digestions success and purification of the desired fragments.

Observations & Results:
The gel did not feature fragments of the expected size.Obviously, something was wrong with the digestion.

V07_02_2 Preparation of over night cultures

Experiment:
In order to gain more plasmid material of the flhDC-promoter constructs over night cultured were prepared for a subsequent plasmid isolation.
20E - #2
20G - #1
2G - #1
20I - #2
18M - #2
18O - #2
18K - #1
18C - #2

V06_12

Preparation of over night cultures

Experiment:
In order to gain further plasmid material over night cultures of one colonie of all eight flhDC-promoter constructs were prepared for subsequent plasmid isolation.
20E - #2
20G - #1
2G - #1
20I - #2
18M - #2
18O - #2
18K - #1
18C - #2

V06_13

V06_13_1 Miniprep of the new flhDC-promoter constructs

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

V06_13_2 Chemical transformartion of pBAD-sfGFP 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.

V06_14

Preparation of over night cultures

Experiment:
Over night cultures of the pBAD-sfGFP clones were prepared in order to isolate the plasmids.

V06_15

Miniprep of pBAD-sfGFP

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

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@@ Line 528: / Line 528: @@
 <td width="900" bordercolor="black" valign="top">
-<h2><b>V06_11 </b></h2><br>
+<h2><b>V07_02 </b></h2><br>
-<b>V06_11_1 Miniprep of the new <i>flhDC</i>-promoter constructs</b><br>
+<b>V07_02_1 Preparative double digestion of 20E-<i>flhDC</i>, 2G-<i>flhDC</i>, 18O-<i>flhDC</i> and pSB1C3</b><br>
+<ul>
-         <ul>
+<li>Experiment:  <br>
-                 <li>Experiment:  <br>
+In order to clone the <i>flhDC</i>-promoter constructs intro pSB1C3, all components were digested with EcoRI and PstI according to the protocol. The restrictio products were subsequently separated on a gel in order to verify the digestions success and purification of the desired fragments. </li><br>
-Minipreps were conducted with PeqGOLD MiniPrep Kit (Peqlab) according to the user manual.
+<li>Observations & Results: <br>
+The gel did not feature fragments of the expected size.Obviously, something was wrong with the digestion. </li>
           </ul>
 <br>
+<b>V07_02_2 Preparation of over night cultures</b><br>
-<b>V06_11_2 Chemical retransformartion of the new <i>flhDC</i>-promoter constructs into <i>E. coli</i> (DH10B)</b><br>
           <ul>
                   <li>Experiment:  <br>
-For the chemical retransformation the standard protocol for transformation was followed. The following constructs were transformed into <i>E. coli</i>:<br>
+In order to gain more plasmid material of the <i>flhDC</i>-promoter constructs over night cultured were prepared for a subsequent plasmid isolation.<br>
+E - #2 <br>
 G - #1 <br>
-G - #2 <br>
 G - #1 <br>
-G - #2 <br>
-G - #3 <br>
 I - #2 <br>
-I - #3 <br>
+M - #2 <br>
 O - #2 <br>
-O - #3 <br>
 K - #1 <br>
-K - #2 <br>
+C - #2 <br></li>
-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>
 </table>