Growing OP50 Cultures for Seeding NGMA Plates
Materials
- Fresh LB streak plate of OP50 (no more than 2 weeks old) stored at 4°C
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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.
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6 cm unseeded NGMA plates (test plates)
- Plastic inoculating loops
Protocol
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Label the autoclaved, sterile 250 mL of LB with "OP50", your initials, and the date.
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Inoculate the 250 mL of sterile LB with a single colony of bacteria from the streak plate using an inoculating loop (Drawer 2-3).
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Grow the culture overnight (16-18 hr: 5 pm to 9 am) in the shaking 37°C incubator in Room 330.
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The next day, immediately store the culture in the 4°C (Rm 223).
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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)
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Indicate on the culture flask label when the culture is ready to use for seeding.
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Grow fresh OP50 each week.