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Team:Amsterdam/Notebook/Protocols/Growth Curve
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==Preparation== | ==Preparation== | ||
| - | 1. Autoclave 250 ml Erlenmeyer’s with 100 ml of LB for each measurement. If you use a magnetic stir, make sure it’s inside as well. | + | 1. Autoclave 250 ml Erlenmeyer’s with 100 ml of LB for each measurement. If you use a magnetic stir, make sure it’s inside as well.<br> |
| - | 2. Autoclave LB for the control measurements and dilutions. | + | 2. Autoclave LB for the control measurements and dilutions.<br> |
| - | 3. Heat or cool baths or stoves 4, 8, 15, 21, 25, 30 and 37oC, with the Erlenmeyer’s already in them. | + | 3. Heat or cool baths or stoves 4, 8, 15, 21, 25, 30 and 37oC, with the Erlenmeyer’s already in them.<br> |
4. Inoculate Top10 cells in 3 ml of LB in greinertubes and grow them overnight at 37 oC. Make sure you at least 1 ml of inoculated TOP 10 cells for each measurement. | 4. Inoculate Top10 cells in 3 ml of LB in greinertubes and grow them overnight at 37 oC. Make sure you at least 1 ml of inoculated TOP 10 cells for each measurement. | ||
==Procedure== | ==Procedure== | ||
| - | 1. Mix all the precultures. | + | 1. Mix all the precultures.<br> |
| - | 2. Put 1 ml off this solution in each of the sterile 250 ml Erlenmeyer’s. | + | 2. Put 1 ml off this solution in each of the sterile 250 ml Erlenmeyer’s.<br> |
| - | 3. Make sure it is properly mixed. | + | 3. Make sure it is properly mixed.<br> |
| - | 4. Measure start OD at 600.nm | + | 4. Measure start OD at 600.nm<br> |
| - | 5. Make sure all measurements remain at their respective temperatures. | + | 5. Make sure all measurements remain at their respective temperatures, and aerated.<br> |
| - | 6. Measure the OD at 600nm every half hour. | + | 6. Measure the OD at 600nm every half hour.<br> |
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| + | Note: Start diluting when your OD600nm measurements go above 0.5<br> | ||
| + | Stop measuring when dilutions of 5x are no longer sufficient<br> | ||
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{{:Team:Amsterdam/Footer}} | {{:Team:Amsterdam/Footer}} | ||
Latest revision as of 00:02, 22 September 2011
Growth curve protocol
Preparation
1. Autoclave 250 ml Erlenmeyer’s with 100 ml of LB for each measurement. If you use a magnetic stir, make sure it’s inside as well.
2. Autoclave LB for the control measurements and dilutions.
3. Heat or cool baths or stoves 4, 8, 15, 21, 25, 30 and 37oC, with the Erlenmeyer’s already in them.
4. Inoculate Top10 cells in 3 ml of LB in greinertubes and grow them overnight at 37 oC. Make sure you at least 1 ml of inoculated TOP 10 cells for each measurement.
Procedure
1. Mix all the precultures.
2. Put 1 ml off this solution in each of the sterile 250 ml Erlenmeyer’s.
3. Make sure it is properly mixed.
4. Measure start OD at 600.nm
5. Make sure all measurements remain at their respective temperatures, and aerated.
6. Measure the OD at 600nm every half hour.
Note: Start diluting when your OD600nm measurements go above 0.5
Stop measuring when dilutions of 5x are no longer sufficient
