Team:Goettingen/week11-3

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Revision as of 16:40, 22 September 2012


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V07_10_1 1^st round: Digestion DpnI/BsaI and clean-up

Experiment:
The digestion and subsequent clean-up were carried out according to the protocol of week 10.

Observations and results:
The corresponding gel showed a band of the expected size.

V07_10_2 1^st round: Ligation

Experiment:
The ligation was carried out according to the protocol of week 10 and incubated over night at 16 °C.

V07_11_1 1^st round: Ethanol precipitation of ligated mutated plasmids

Experiment:
The ethanol precipitation was carried out according to protocol, this time with the right volumes!

Observations and results:
The corresponding gel showed a band of the expected size.

V07_11_2 1^st round: Transforamtion of electrocompetent cells wir the mutant plasmid mixture

Experiment:
Electrocompetent cells were prepared according to protocol. We transformed 200 µL of cells with 10 µL of DNA according to the protocol of week 10. Dilution plates and liquid culture was incubated over night at 37 °C, 180 rpm.

V07_12_1 1^st round: Analysis of transformation V07_11

Experiment:
Both the liquid culture and the dilution plates were checked for bacterial growth. 5 mL LB liquid cultures + CM were prepared for each clone on the plates for subsequent sequencing of the plasmids. The cultures were incubated over night at 37 °C and 180 rpm.

Observations and results:
Both the liquid culture and the dilution plates were positive for bacterial growth. Plates showed the following results:
10⁴ 10 clones
10⁵ 2 clones
10⁶ 0 clones

V07_12_2 1^st round: Miniprep of liquid culture V07_11

Experiment:
We used 4 x 5 mL liquid culture split to 4 columns for plasmid preparation with the peqGOLD Plasmid Miniprep Kit (PeqLab).

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 <b>V07_12_2 1<sup>st</sup> round: Miniprep of liquid culture V07_11</b><br>
 <ul>
-<li>Experiment: <br>We used 4 x 5 mL liquid culture onto 4 columns for plasmid preparation with the peqGOLD Plasmid Miniprep Kit (PeqLab).
+<li>Experiment: <br>We used 4 x 5 mL liquid culture split to 4 columns for plasmid preparation with the peqGOLD Plasmid Miniprep Kit (PeqLab).
 </li>
 </ul>