<li>Observations and results: <br>Both the liquid culture and the dilution plates were positive for bacterial growth. Plates showed the following results:<br>
+
<li>Observations and results: <br>Concentrations varied between 111 and 165 ng/µL.
-
10<sup>4</sup> 10 clones<br>
+
-
10<sup>5</sup> 2 clones<br>
+
-
10<sup>6</sup> 0 clones<br>
+
</li>
</li>
</ul><br>
</ul><br>
-
<b>V07_13_2 1<sup>st</sup> round: Miniprep of liquid culture V07_11</b><br>
V07_10_1 1st 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 1st round: Ligation
Experiment: The ligation was carried out according to the protocol of week 10 and incubated over night at 16 °C.
V07_11
V07_11_1 1st 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 1st 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
V07_12_1 1st 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 1st 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).
V07_13
V07_13_1 1st round: Miniprep of clones 1-12 V07_12
Experiment: Plasmids from the 1st mutation round were prepped using the peqGOLD Plasmid Miniprep Kit (PeqLab) following the user manual. DNA concentrations were determined for subsequent sequencing.
Observations and results: Concentrations varied between 111 and 165 ng/µL.
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