From 2012.igem.org
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I. Check overnight cultures for growth
- As of ~2:30 PM when I got in, both cultures were cloudy corresponding to saturation
II. Make 60% glycerol stock from 100% stock
- Dispense 1.6 mL of sterile dH2O into a 15 mL conical tube
- Add 2.4 mL of sterile 100% glycerol, making sure to wait to make sure the viscous fluid is completely dispensed
- Mix thoroughly by vortex
III. Make 15% Glycerol Stocks (6 total) from Successful Overnight Cultures
- Transfer 600 uL of each liquid culture to each microcentrifuge tube (x3 each)
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- Add 200 uL of 60% glycerol to each tube (x3 each)
- Vortex to mix thoroughly
- Store tubes in the -80 oC freezer
- Discard any remaining liquid culture from above cell line
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