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Team:Paris Saclay/Notebook/July/23
From 2014.igem.org
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==Lab work== | ==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 | ||
| + | # Add equal parts plasmid DNA and ethanol 100% in a 1.5 microcentrifuge tube. | ||
| + | # 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) | ||
| + | # Centrifuge for 10 minutes at 4°C and at 11000g. | ||
| + | # Discard supernatant. | ||
| + | # Add 1ml of ethanol 70%. | ||
| + | # Repeat step 3. | ||
| + | # 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. | ||
| + | # (tomorrow) Resuspend pellet in 30μl H<sub>2</sub>O milliQ. | ||
==Reunion== | ==Reunion== | ||
Revision as of 21:27, 23 July 2014
Contents |
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
- Add equal parts plasmid DNA and ethanol 100% in a 1.5 microcentrifuge tube.
- 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)
- Centrifuge for 10 minutes at 4°C and at 11000g.
- Discard supernatant.
- Add 1ml of ethanol 70%.
- Repeat step 3.
- 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.
- (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.
