User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2013/02/05

Experimental Biological Chemistry II

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Purpose Grind Ag+ Clay into powder for X-ray spectroscopic analysis. Grow DH5α-T1 bacteria cells and make two glycerol stocks of it. Procedure 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.