Team:Goettingen/week9-2

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Revision as of 11:19, 15 September 2012

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V06_25_1 Chemical transformartion of the BioBrick plasmid pSB1C3 into E. coli (DH10B)

Experiment:
In order to gain further plasmid material of the new vector pSB1C3 was transformed into E. coli(DH10B) as described in the protocol.
Observations & Results:
The transformation failed. Only the positive control featured colony formation. We found out that accidentally ampicillin containing plates were used whereas the plasmid obtains a chloramphenicol resistance. Thus, the out come is not surprising.

V06_25_2 Preparation of over night cultures

Experiment:
Since we received a new E. coli (DH10B) strain, over night cultures were prepared in order to produce new competent cells.The following protocol was considered.

V06_26_1 Preparation of chemically competent cells of the new E.coli (DH10B) strain

Experiment:
The preparation of the competent cells was performed according to the following protocol.
Observations & Results:
The performance of a test transformation proved the functionality of the new competent cells.

V06_26_2 Preparation of over night cultures

Experiment:
In order to test different E. coli strains competent cells of the recently received strains BL21, DH5alpha and XL1 Blue should be produced. On that account over night cultures of al strains were prepared.

V06_27_1 Preparation of chemically competent cells of the new E.coli strains BL21, DH5alpha and XL1 Blue

Experiment:
The preparation of the competent cells was performed according to the following protocol.

V06_27_2 Chemical transformation of flhDC-promoter constructs E.coli ( DH10B, BL21, DH5alpha and XL1 Blue)

Experiment:
For the chemical transformation the standard protocol was considered. The following constructs were transferred into the cells:
18O-fhlDC
18M-fhlDC
18C-fhlDC
Observations & Results:
The transformation of the DH10B cells functioned very well. Numerous colonies could be found at all plates except for the negative control. The other three strains did not lead to such good results. Here, now or low bacterial growth could be observed. Since the competent cells of DH10B worked so well, we suggested that the other strains were just too fresh, so the transformation should be repeated.

Preparation of over night cultures

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 </ul>
 <br>
-<b>V06_27_2 Chemical transformation of <i>flhDC</i> under control of three distinct promoters into <i>E.coli</i> (BL21, DH5alpha and XL1 Blue)</b><br>
+<b>V06_27_2 Chemical transformation of <i>flhDC</i>-promoter constructs <i>E.coli</i> ( DH10B, BL21, DH5alpha and XL1 Blue)</b><br>
 <ul>
 <li>Experiment:  <br>
-In order to test different <i>E. coli</i> strains competent cells of the recently received strains BL21, DH5alpha and XL1 blue should be produced. On that account over night cultures of al strains were prepared. </li>
+For the chemical transformation the standard protocol was considered. The following constructs were transferred into the cells: <br>
+O-<i>fhlDC</i> <br>
+M-<i>fhlDC</i> <br>
+C-<i>fhlDC</i> <br></li>
+<li>Observations & Results: <br>
+The transformation of the DH10B cells functioned very well. Numerous colonies could be found at all plates except for the negative control. The other three strains did not lead to such good results. Here, now or low bacterial growth could be observed. Since the competent cells of DH10B worked so well, we suggested that the other strains were just too fresh, so the transformation should be repeated.
+</li>
 </ul><br>