Team:Goettingen/week9-2

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Revision as of 10:51, 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 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

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.
Observations & Results:
The performance of a test transformation proved the functionality of the new competent cells.

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

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 <ul>
 <li>Experiment: <br>
-Since we received a new <i>E. coli</i> (DH10B) strain, over night cultures were prepared in order to produce new competent cells.<br>
+Since we received a new <i>E. coli</i> (DH10B) strain, over night cultures were prepared in order to produce new competent cells.The following protocol was considered.<br>
 <li>Observations & Results: <br>
 The performance of a test transformation proved the functionality of the new competent cells.