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

Experimental Biological Chemistry II

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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.