User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2013/01/30

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Experimental Biological Chemistry II Main project page
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Purpose

  • The purpose today is to autoclave LB medium to prepare for cell culture.
  • Also, Ag+ incorporated clay is going to be filtered with water and ethanol to isolate clay from water/ethanol solution.

Procedure

  • 50mL LB media was made using the following way:
  0.50g Bacto-Trypsin
  0.25g Bacto-Yeast Extract
  0.50g Sodium Chloride
  50mL of distilled water
  Note: LB media obtained from Genova® with 2:1:2 proportion was used instead. 1.25g of LB powder was weighted out.
  • Each of the 50mL LB media was placed in two 125mL glass bottles and four 250mL Erlenmeyer flask.
  • 500mL of Agar was made using the following in a 1000mL Erlenmeyer flask:
  12.5g of LB media powder
  500mL of distilled water
  7.5g of Trypsin Soy Agar
  • The Erlenmeyer flask obtaining LB media or Agar were covered with aluminum foil, the bottles containing LB were capped loosely. All media were autoclaved under liquid cycle.
  • The clay+AgNO3 mixture was taken off hot plate.
  • A filter funnel, attached to 500mL Erlenmeyer flask then attached to a pump, is covered with a paper filter. Surface of paper filter was wetted and attached onto filter funnel through adding water.
  • The clay+AgNO3 mixture was then transfered into the filter funnel with suction applied.
  • Aluminum foil was used to cover the funnel to avoid silver to interact with light.
  • After autoclave, the LB media were placed on counter without removing the aluminum foil. The Agar were cooled to about 50°C, until the wall of Erlenmeyer flask can be touched by hand. Approximately 20mL of the Agar were poured into each of the 20 separate petri dishes.

Results

  • The clay+AgNO3 mixture when taken off the hot plate appeared black. However, during filtration, the filtered liquid solution remained clear in color. However, the filtered liquid solution turned dark and obtained a darker red color after sitting on the counter. The filtered liquid solution was analyzed by Dr. Fox, who later concluded that silver nanoparticles were found in filtrate.



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