- Remake LB+agar plates
- Make glycerol stock of DH10B and DH5α-T1 cells
- Test absorbance of cells in culture grew last night for cell concentration
- Because agar plates made on 2013/01/31 liquified, new plates were made.
- 12.5g of LB media powder were weighted and placed in a 750mL Erlenmeyer flask.
- 7.5g of Agar were weighted out and placed in the same 750mL Erlenmeyer flask.
- 500mL distilled water was measured out and placed in the same 750mL Erlenmeyer flask.
- The solution was autoclaved per liquid cycle.
- After autoclave, the agar were placed in room temperature to cool down to 60°C.
- Approximately 25mL of agar were poured into each petri dish.
- The agar plates were left in room temperature under flame to solidify.
- The agar plates were stored in fidge for future use.
- Glycerol stock of the two cell lines: DH10B and DH5α-T1 cells were made for future use.
- Two 5mL Gyro® Tubes were obtained, one for DH10B cells and another for DH5α-T1 cells.
- 150mL of autoclaved 60% glycerol in water was pipetted into the Gyro® Tubes prior.
- 850mL of DH10B cells, and 850mL of DH5α-T1 cells were pipetted into each of Gyro® Tubes, respectively.
- Each cell culture containing the cell and glycerol were vortexed until mix.
- Cells were placed in -80°C for storage.
- Absorbance of cells were taken using UV-vis spectroscopy to measure the concentration of cells that had been grown over night.
- UV-vis spectrometer was set under photometric method, with wavelength set at 650nm.
- A distilled water sample placed in plastic cuvette was placed into the spectrometer to eliminate background noise.
- 2mL of DH10B and DH5α-T1 cells were individually placed into a plastic cuvette into the spectrometer to measure absorbance.
- The absorbance for each cell after 15 hours of growth is shown in table below: