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From 2014.igem.org

Contents 1 Wednesday 23rd July 1.1 Lab work 1.1.1 1 - p cola part 2 1.2 Reunion

Wednesday 23rd July

Lab work

1 - p cola part 2

by Sean

p cola DNA was extracted in several instances yesterday. Today we attempt to collect all of the DNA and precipitate it.

Protocol

1. Add equal parts plasmid DNA and ethanol 100% in a 1.5 microcentrifuge tube.
2. If total volume is V, then add V/5 of sodium acetate (initial concentration of NaAc is 3M; we want a final concentration of .3M). Numerical application: Total volume of p cola was 150μl. Hence 150 μl of ethanol 100% was added, as well as 30μl of NaAc 3M. Incubate at -20°C for 30 minutes. (5:20pm - 5:50pm in our case)
3. Centrifuge for 10 minutes at 4°C and at 11000g.
4. Discard supernatant.
5. Add 1ml of ethanol 70%.
6. Repeat step 3.
7. Discard supernatant. Make tube interior as dry as possible. Tip: use a paper towel on which to lightly tap the rim of the tube. Dry pellet in oven at 37°C. Note: we decided to leave the tube at room temperature and continue tomorrow instead.
8. (tomorrow) Resuspend pellet in 30μl H₂O milliQ.

Reunion

We talk about the organisation of the wiki's notebook, the different parts of our project and the ethic points.

Members there:

• Instructors and advisors: Solenne and Sylvie.
• Students: Arnaud, Eugene, Fabio, Juliette, Leila, Pierre, Romain, Sean and Terry.

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