Purpose:
To isolate the plasmid DNA from the rest of the bacteria for colonies containing LuxPQ on TOPO II Blunt and LuxPQ BBk on psB1AC3.
Reference: The user manual and the Sigma Aldrich site
Kit: MP70 GeneElute Plasmid mini-prep kit. Note that it had been previously used, which may be a problem for contamination.
Materials and methods:
Procedure was carried out in accordance with our Plasmid Isolation protocol. When complete, we stored our isolated plasmids in the freezer for concentration measurement and content analysis over the next 2 days.
Results:
We were left with tubes containing the described plasmids. Quantifiable details on their concentrations and actual contents are outlined on the May 27 and May 28 entries.
DNA concentration measurements with the Nanodrop spectrophotometer
Purpose:
To measure the DNA concentration of LuxPQ plasmids that were isolated in the May 26 miniprep
Materials and Methods:
The DNA measurement area was cleaned with isopropanol and KimWipes to clear off any trace residue from previous uses. The sample wavelength was set to 260 nm. A blank measurement was made with 1 uL of ddHOH. 1 uL of the sample in question was added to the Nanodrop measurement area. The measurement was made; the sample area was wiped down; and a new sample was measured.
Results:
| Plasmid | 260/280 | 260/230 | Concentration [ng/μL]
|
| LuxPQ TOPO C1 | 1.83 | 1.86 | 338.2
|
| LuxPQ TOPO C2 | 1.81 | 1.87 | 284.2
|
| LuxPQ BBK C1 | 1.90 | 2.34 | 295.5
|
| LuxPQ BBK C2 | 1.90 | 2.29 | 313.2
|
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