Making a long term stock of bacteria
From OpenWetWare
(Difference between revisions)
m |
|||
| Line 3: | Line 3: | ||
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 04:34, 28 June 2012
| back to protocols | ||
Contents |
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
