| Experimental Biological Chemistry II
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- Grind Ag+ Clay into powder for X-ray spectroscopic analysis.
- Grow DH5α-T1 bacteria cells and make two glycerol stocks of it.
- Procedure for grinding of Ag+ Clay into powder form is recorded in Melissa's Notebook.
- Growing of DH5α-T1 cells
- One shot of DH5α-T1 cells was taken out from -87°C fridge.
- Cells were thawed in room temperature
- 3mL of autoclaved LB media was pipetted into a test tube
- Inoculation loop was used to obtain a small portion of cells. The cells were then dipped into the 3mL LB media.
- The cell culture was then placed in orbital shaker for 2 hours, at 37°C, with shaking at 230rpm.
- After two hours, the culture appeared clear, indicating no cell growth, an alternative approach were taken.
- A different cell line was used for cell amplification, and the procedure for DH5α-T1 cell amplification was altered then repeated.
- DH10B cells were taken from -87°C fridge.
- The cell were let thaw on ice.
- In two clean 15mL falcon tubes, 3mL autoclaved LB media were added.
- In one falcon tube, 10uL DH10B cells were added.
- In another falcon tube, 10uL DH5α-T1 cells were added.
- The two falcon tubes were placed in orbital shaker for 2 hours, at 37°C, with shaking at 230rpm.
- After 2 hours of incubation, the both cultures appeared clear. As a result, both cultures were placed in incubator horizontally to maximize shaking and left in shaker under same conditions for over night.