Making a long term stock of bacteria: Difference between revisions
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Whenever you successfully transform a bacterial culture with a plasmid or whenever you obtain a new bacterial strain you will want to make a long term stock of that bacteria. Bacteria can be stored for months and years if they are stored at -80C and in a high percentage of glycerol. | Whenever you successfully transform a bacterial culture with a plasmid or whenever you obtain a new bacterial strain you will want to make a long term stock of that bacteria. Bacteria can be stored for months and years if they are stored at -80C and in a high percentage of glycerol. | ||
== | ==Materials== | ||
*40% glycerol solution | |||
* Day/overnight culture | |||
*Cryogenic vials/1.5mL microfuge tube | |||
==Method== | |||
*Add 1 ml of 40% glycerol in H<sub>2</sub>O 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. | |||
==Specific Protocols== | |||
*[[Endy:Making a long term stock of bacteria]] | *[[Endy:Making a long term stock of bacteria]] | ||
*[[Endy:Retrieving a Registry glycerol]] | *[[Endy:Retrieving a Registry glycerol]] | ||
*[[-80_Glycerol_Stocks]] | |||
*[[Alm:Glycerol_stocks]] | |||
*[[McClean:E._coli_Glycerol_Stocks]] | |||
*[[Cfrench:glycerol]] | |||
*[[Smolke:Protocols/Freezer_stocks]] | |||
==Notes== | ==Notes== | ||
Revision as of 01:34, 28 June 2012
| back to protocols | ||
Introduction
Whenever you successfully transform a bacterial culture with a plasmid or whenever you obtain a new bacterial strain you will want to make a long term stock of that bacteria. Bacteria can be stored for months and years if they are stored at -80C and in a high percentage of glycerol.
Materials
- 40% glycerol solution
- Day/overnight culture
- Cryogenic vials/1.5mL microfuge tube
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.
Specific Protocols
- Endy:Making a long term stock of bacteria
- Endy:Retrieving a Registry glycerol
- -80_Glycerol_Stocks
- Alm:Glycerol_stocks
- McClean:E._coli_Glycerol_Stocks
- Cfrench:glycerol
- Smolke:Protocols/Freezer_stocks
