Team:NDC-HighRiverAB/Notebook
NOTEBOOK
Wednesday, July 4, 2018
Today we made sterile LB and some new agar plates (5 with amp, 1 without + some extra plates that should have had amp but didn't.) Ampicillin was added at a ration of 1 uL per mL of agar liquid.
We autoclaved some waste and some pipette tips.
We received competent cells from fredsense to use for a transformation with our NDC 001-1 in the PET21a plasmid.
We had an issue with rehydrating the DNA. We added 15 uL of distilled water to 4 ug of plasmid, but after centrifuging and stirring it seemed like there was very little DNA solution, so we added an additional 10 uL of water before proceeding with the transformation. We let this sit for 7 minutes before moving forward with the transformation.
Because of needing to rehydrate the DNA with additional water, the competent cells were thawed on ice, distributed into tubes, and then put back on ice before the DNA was added.
We followed the transformation procedure from the Synthetic Biology Workshop Protocol Manual (University of Lethbridge 2016).
The following plates were made:
| Plate | Competent Cells | Transformed with Plasmid | Ampicillin |
|---|---|---|---|
| Control | X | ||
| Control | X | X | |
| Plate 1 | X | X | X |
| Plate 2 | X | X | X |
| Plate 3 | X | X | X |
| Plate 4 | X | X | X |
| Plate 5 | X | X | X |