Team:Macquarie Australia/Notebook
- We went through lab safety induction to ensure safety procedures were followed at all times in the lab
- We learnt how to perform digestion, ligation, transformation and how to make and run agarose gels
- We used digestion to screen the Photosystem II (PSII) parts DCA, psbELJTB and psbMZtlwkOPQR. psbELJTB and psbMZtlwkOPQR showed successful digestion and correct sizes. DCA showed signs of contamination in all samples.
- We decided to focus of the Chlorophyll plasmid rather than PS II. We proceeded with digestion and ligation of ChlD with ChlI2 as well as DRV1 with ChlG, both into Kanamycin backbones, following the 3A assembly protocol.
- Renee, Areti and Karl also began the interlab study:
- Made LB media + cam plates
- Transformed Εscherichia coli DH5α cells with the interlab testing devices.Transformation was performed twice, due to poor colony growth in the LB + cam media at the first round of transformations.
- We transformed the DVR1-ChlG and ChlD-ChlI2 ligation samples into competent DH5α E.coli cells. DVR1-ChlG had successful transformants which were then incubated in LB broth + Kan. ChlD-ChlI2 gave an unsuccessful transformation, so the process was repeated successfully this time. The samples were then liquid cultured.
- Interlab experiments were completed this week:
- Calibrations 1, 2 and 3 were completed early on in the week
- Cell measurements (Abs600 and fluorescence) were taken as per the interlab protocol
- Colony forming units per OD600 = 0.1 of the negative and positive control devices was also determined (in triplicates)
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