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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Experimental Biological Chemistry II
