Making long term stocks | Transformation | [http://microscopy.berkeley.edu/Resources/instruction/buffers.html Buffer recipes] | Colony PCR

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Making long term stocks in glycerol

(From http://openwetware.org/wiki/Endy:Making_a_long_term_stock_of_bacteria)

Materials

• 40% glycerol solution
• Cryogenic vials

Method

• Add 1 ml of 40% glycerol in H₂O to a cryogenic vial.
• Add 1 ml sample from the culture of bacteria to be stored.
• Gently vortex the cryogenic vial to ensure the culture and glycerol is well-mixed.
  • Alternatively, pipet to mix.
• Use a tough spot to put the name of the strain or some useful identifier on the top of the vial.
• On the side of the vial list all relevant information - part, vector, strain, date, researcher, etc.
• Store in a freezer box in a -80C freezer. Remember to record where the vial is stored for fast retrieval later.

Notes

• While it is possible to make a long term stock from cells in stationary phase, ideally your culture should be in logarithmic growth phase.