Add one ml overnight culture to a cryotube from step 1
Store at -80 °C
PCR
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).
Casting of gels and gel electrophoresis (remember to include EtBr)
A
A
S
Preparation of Cam stock solution
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u
i
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
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
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.