Team:Goettingen/week8-2

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Revision as of 10:01, 15 September 2012

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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 was not successful. No colonies could be observed at the plates. Since this vector has a chloramphenicol resistance instead of ampicillin as the usually used pUC18 we suggest that the antibiotics concentration in the agar was simply to high.

V06_19_1 Repetition of the chemical transformartion of the BioBrick plasmid pSB1C3 into E. coli (DH10B)

Experiment:
Minipreps were conducted with PeqGOLD MiniPrep Kit (Peqlab) according to the user manual.
Observations & Results:
The transformation was successful.Due to the possesion of a gene encoding for RFP the developed colonied featured a red color. On the negative control no colonied were observed.

V06_19_2 Repetition of the chemical transformartion of the araC-promoter into E. coli (DH10B)

Experiment:
Since the last time the fact that the araC-promoter I0500 (14N) is inducible by ITPG was not considered, this time the substance was added to the LB-Medium (end concentration 100 µM). This is the only deogation from the standard transformation protocol.
Observations & Results:
The transformation was again not successful. Only the positive control showed any growth.

Preparation of over night cultures

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

Miniprep of pBAD-sfGFP

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

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

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 </li></ul>
 <br>
-<b>V06_19_2 Chemical transformartion of pBAD-<i>sfGFP</i> into E. coli (DH10B)</b><br>
+<b>V06_19_2 Repetition of the chemical transformartion of the <i>araC</i>-promoter into <i>E. coli</i> (DH10B) </b><br>
 <ul>
 <li>Experiment:  <br>
-The chemical transformation was conducted as described in the protocol, however, a lower chloramphenicol concentration was used than before.<br>
+ Since the last time the fact that the <i>araC</i>-promoter I0500 (14N) is inducible by ITPG was not considered, this time the substance was added to the LB-Medium (end concentration 100 µM). This is the only deogation from the standard transformation protocol. <br>
 <li>Observations & Results: <br>
-The transformation was successful since all plates showed numeours colonoes except the negative control.</li>
+The transformation was again not successful. Only the positive control showed any growth.  </li>
 </ul><br>