Revision as of 21:28, 23 July 2014

Wednesday 23rd July

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.

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.

@@ Line 15: / Line 15: @@
 # 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.
+# 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. <span style="color:red">Note: we decided to leave the tube at room temperature and continue tomorrow instead.</span>
 # (tomorrow) Resuspend pellet in 30μl H<sub>2</sub>O milliQ.