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

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.