From 2014.igem.org
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- | <ol> | + | <ul> |
- | <li>Add one ml sterile 50% glycerol to a cryotube.</li> | + | <li>Make a mastermix of the reagents that are common for all reactions. Make enough for 0,5 extra reaction. Add reagents in the order they are listed above.</li> |
- | <li>Add one ml overnight culture to a cryotube from step 1</li> | + | <li>The polymerase stock must only be taken out of the freezer when needed and must be put back immediately after use.</li> |
- | <li>Store at -80 °C</li> | + | <li>Remember to mix thoroughly before aliquotting to separate PCR tubes, and try not to make any bubbles. Reverse pipetting works well for aliquoting.</li> |
- | </ol> | + | <li>Mark the PCR tubes on lid and side and place in thermocycler.</li> |
| + | <li>The annealing temperature should match the primer melting temperatures. This can be hard as Tm predictors tend to disagree.</li> |
| + | <li>Elongation time should be AT LEAST 30 seconds per kb of the longest expected fragment (for the X7 polymerase).</li> |
| + | </ul> |
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Revision as of 14:03, 1 August 2014
Protocols
Glycerol stock-preservation |
- Add one ml sterile 50% glycerol to a cryotube.
- Add one ml overnight culture to a cryotube from step 1
- Store at -80 °C
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PCR |
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- Make a mastermix of the reagents that are common for all reactions. Make enough for 0,5 extra reaction. Add reagents in the order they are listed above.
- The polymerase stock must only be taken out of the freezer when needed and must be put back immediately after use.
- Remember to mix thoroughly before aliquotting to separate PCR tubes, and try not to make any bubbles. Reverse pipetting works well for aliquoting.
- Mark the PCR tubes on lid and side and place in thermocycler.
- The annealing temperature should match the primer melting temperatures. This can be hard as Tm predictors tend to disagree.
- Elongation time should be AT LEAST 30 seconds per kb of the longest expected fragment (for the X7 polymerase).
|
Glycerol stock-preservation |
- Add one ml sterile 50% glycerol to a cryotube.
- Add one ml overnight culture to a cryotube from step 1
- Store at -80 °C
|
Glycerol stock-preservation |
- Add one ml sterile 50% glycerol to a cryotube.
- Add one ml overnight culture to a cryotube from step 1
- Store at -80 °C
|
Glycerol stock-preservation |
- Add one ml sterile 50% glycerol to a cryotube.
- Add one ml overnight culture to a cryotube from step 1
- Store at -80 °C
|
Glycerol stock-preservation |
- Add one ml sterile 50% glycerol to a cryotube.
- Add one ml overnight culture to a cryotube from step 1
- Store at -80 °C
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Rasmus Bech |
Motivation
I joined iGEM in order to take part in a project where the students have more control and responsibility than other projects at DTU. It was also a motivation being part of a team, as large projects at DTU most often are individual.
Major contributions
How do you prefer spinach?
I prefer spinach in a salad with strawberry, red onion, pine nuts, feta and a dressing with Balsamico vinegar, oil, sugar, soy sauce and tabasco.
Thor Bech Johannesen |
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Motivation
As a student at DTU you get to do plenty of project work, however it is mostly carried out individually or in a small group, with very specific goals and a high degree of supervision. iGEM offers the opportunity to work on a much larger project with a high degree of autonomy, where everything from the overall goals and strategy to design of experiments is left for the students to manage. This provides a great opportunity to experience the challenges of actual research projects
Major contributions
How do you prefer spinach?
Luminescent and far away from my taste buds.
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Overview |
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