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

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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.



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