User:Melissa Novy/Notebook/CHEM-572/2013/01/30

Objectives

• Make 6 sets of 50-mL LB media.
• Make 500 mL agar solution.
• Autoclave LB media and agar solution.
• Make 20 agar plates.
• Filter and dry LMT-95Ag, made on 2013/01/29.

LB Media

• Protocol
  1. Obtain 6 clean, dry 250-mL Erlenmeyer flasks.
  2. Place 1.25 g of LB medium, obtained from Teknova, into each flask.
  3. Fill each flask with 50 mL deionized H₂O.
  4. Cap each flask with aluminum foil and swirl until the LB dissolves.
  5. Autoclave the flasks according to the AU Biomaterials Design Lab protocol.

• Note that the protocol was adapted from the OpenWetWare protocol.

Agar Solution and Agar Plates

• Protocol
  1. Obtain a clean, dry 1000-mL Erlenmeyer flask.
  2. Place 12.5 g of LB medium, obtained from Teknova, into the flask.
  3. Fill the flask with 500 mL deionized H₂O.
  4. Swirl the flask to dissolve the LB, then add 7.5 g Trypticase Soy Agar, obtained from Aldrich.
  5. Cap the flask with aluminum foil and autoclave according to the AU Biomaterials Design Lab protocol.
  6. Obtain 20 sterile 10-cm Petri dishes.
  7. Fill each Petri dish with 25 mL of autoclaved agar solution.

Filter and Dry LMT-95Ag

• LMT-95Ag was covered with aluminum foil and left to stir for 24 h. It was then filtered with Whatman 41 filter paper and a clean, dry filter flask and funnel. It was washed with 10 mL of (50:50) H₂O-EtOH.
• LMT-95Ag was then scraped from the filter paper with a spatula and placed in an aluminum dish loosely capped with foil in an 80°C oven to dry overnight.
• The filtrate was reserved for further testing for sodium.

• Observations
  • LMT-95Ag turned from opaque white in color to opaque dark gray. This indicates possible oxidation of Ag.
  • The filtrate was clear and colorless, but became transparent red-gray in color after about 30 min. The filtrate was subsequently covered in foil.