Team:Arizona State/Lab/Protocols/Cultures

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.