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Liquid Culture:
- Pick single colony from plate with pipet tip.
- Drop tip in tube of ~7 mL LB media (with selected antibiotic if necessary).
- Shake overnight at 37C.
Plating Cells:
From solution/liquid culture:
- Apply 100-200 μl cell solution to LB agar plate containing appropriate antibiotic.
- Place spreader in 95% ethanol, remove and sterilize in flame.
- Using sterilized spreader, gently spread solution around plate.
- Store upside down at 37C overnight to grow cells.
- Store at 4C long-term.
Streak plating:
- Place inoculation loop in 95% ethanol, remove and sterilize in flame.
- Cool inoculation loop by pressing into a section of agar in used plate.
- Pick single colony of cells from original plate.
- Streak colony across new plate.
- Store upside down at 37C overnight to grow cells.
- Store at 4C long-term.
Glycerol Stock:
- Prepare solution of 40% glycerol by volume.
- Add 750 μl cell solution and 750 μl glycerol solution (or 1:1 mix) to Eppendorf tube.
- Store at -80C.
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