Growing OP50 Cultures for Seeding NGMA Plates

Materials

  • Fresh LB streak plate of OP50 (no more than 2 weeks old) stored at 4°C
  • 250 mL LB in 500 mL flask(s) (usually every Monday)

    Note: This may already be prepared, but if not, prepare by autoclaving LB in flasks.

  • 6 cm unseeded NGMA plates (test plates)

  • Plastic inoculating loops

Protocol

  1. Label the autoclaved, sterile 250 mL of LB with "OP50", your initials, and the date.

  2. Inoculate the 250 mL of sterile LB with a single colony of bacteria from the streak plate using an inoculating loop (Drawer 2-3).

  3. Grow the culture overnight (16-18 hr: 5 pm to 9 am) in the shaking 37°C incubator in Room 330.

  4. The next day, immediately store the culture in the 4°C (Rm 223).

  5. Spot 6 cm NGM test plates (2 per culture) with 100 μL of culture and place at 37°C overnight to test for normal growth and no contamination. (optional)

  6. Indicate on the culture flask label when the culture is ready to use for seeding.

  7. Grow fresh OP50 each week.