Team:EPF-Lausanne/Protocol/Gel

From 2012.igem.org

(Difference between revisions)
Line 16: Line 16:
# Microwave, at 7, the bottle (no cap!) until it boils.
# Microwave, at 7, the bottle (no cap!) until it boils.
# Carefully remove bottle (can be super heated!) and check for the total absence of particles. Microwave again if needed.
# Carefully remove bottle (can be super heated!) and check for the total absence of particles. Microwave again if needed.
-
# Prepare a gel box and fill it up with the agarose solution (maybe not the whole solution is needed).
+
# Prepare a gel box, with comb, and fill it up with the agarose solution (maybe not the whole solution is needed).
# Add 0.05 µl per ml of gel in the box of Red Gel (it's in the iGEM drawer) and stirr until disolved.
# Add 0.05 µl per ml of gel in the box of Red Gel (it's in the iGEM drawer) and stirr until disolved.
-
# UNFINISHED
+
# Wait until cold and solidified.
 +
# Carefully remove comb.
 +
# Place the box in the voltage generator.
 +
# Cover the gel with buffer. (What buffer?)
 +
# Add blue dye to the DNA samples and ladder marker. (Which dye? How much?)
 +
# Inject 20 µl of ladder marker in the first well.
 +
# Inject 20 µl of each DNA solution in the other wells.
 +
# Ready to run.
 +
UNFINISHED
{{:Team:EPF-Lausanne/Template/ProtocolFooter}}
{{:Team:EPF-Lausanne/Template/ProtocolFooter}}
<noinclude>{{:Team:EPF-Lausanne/Template/Footer}}</noinclude>
<noinclude>{{:Team:EPF-Lausanne/Template/Footer}}</noinclude>

Revision as of 01:07, 2 July 2012

Protocol: Gel electrophoresis


Agarose concentration depends on the size of the DNA to be run. We will mostly use 1%. VOL is the desired volume of gel in ml:

  1. Add 0.01*VOL g of agarose to a clean glass bottle.
  2. Pour VOL/50 ml of 50xTAE in a graduated cylinder. Fill up to VOL ml with di water.
  3. Add the resulting VOL ml of 1xTAE to the glass bottle with agarose.
  4. Microwave, at 7, the bottle (no cap!) until it boils.
  5. Carefully remove bottle (can be super heated!) and check for the total absence of particles. Microwave again if needed.
  6. Prepare a gel box, with comb, and fill it up with the agarose solution (maybe not the whole solution is needed).
  7. Add 0.05 µl per ml of gel in the box of Red Gel (it's in the iGEM drawer) and stirr until disolved.
  8. Wait until cold and solidified.
  9. Carefully remove comb.
  10. Place the box in the voltage generator.
  11. Cover the gel with buffer. (What buffer?)
  12. Add blue dye to the DNA samples and ladder marker. (Which dye? How much?)
  13. Inject 20 µl of ladder marker in the first well.
  14. Inject 20 µl of each DNA solution in the other wells.
  15. Ready to run.

UNFINISHED